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ISSN 0840-8440 










TD Environmental research 

j_72 5 technology transfer conference 

Q57 1988 proceedings / 

1988 20307 
vol. 2 

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ISSN 0840-8440 


November 28 and 29, 1 988 
Royal York Hotel 
Toronto, Ontario 


Sponsored by 
Research and Technology Branch 
Environment Ontario 
Ontario, Canada 

( 6^M^vrKS ) 


Environment Ontario holds its annual Technology Transfer 
Conference to report and publicize the progress made on 
Ministry-funded projects. These studies are carried out in 
Ontario Universities and by private research organlzationa 
and companies. 

The papers presented at Technology Transfer Conference 1988 

are published in five volumes of conference Proceedings 
corresponding to the following sessions; 






This volume is comprised of presentations given during 
Session B of the conference. 

For reference purposes, indices for sessions A,C,D and E 
may be found at the back of this volume, listed in alpha- 
numeric order- 

For further information on any of the papers, please 
contact either the authors or the Research and Technology 
Branch at (-Jie) 323-4574 or 323-4573. 

Aclcnowl edgementa 

The Conference Committee would like to thank the authors 
for their valuable contributions to environmental research 
in Ontario. 


The views and ideas expressed in these papers are those of 
the authors and do not necessarily reflect the views and 
policies of Environment Ontario, nor does mention of trade 
names or commercial products constitute endorsement or 
recommendation for use. 


Keynote Papers 

KsynotB Paper l! Sci«nce-ba««d Innovation and 
Proaparlty Within "Sustalnabla Devolopmant"; 
J. Fraser Mustard, The Canadian Institute for 
Advance Research, Toronto, Ontario. 

K«ynote Paper II: Darlving Banefita from 
Environmental Research; Stuart Smith, 
RockcliCfe Research and Technology Inc., Ottawa 

Environment Ontario Paper 

Mat«r Quality Reaearch: Current Statue and Future 
Raaearah Naede; C. Schenk, Water Resources Branch, 
Environment Ontario, 

Abstract Page 

Oral Presentations 

B1 Aquatic Biology in the New Regulatory Framework K. 13 

Day. National Water Reseach Institute, Burlington. 

B2 HypothesisTosting InAquaticToxicology; QSAR 15 

Helationshlps and Simple Kinetic Model ling L. S. 
McCarty". University of Waterloo. Waterloo. Ontario, 
G.W, OzburnandA.D. Smith. Lakehead University. 
Thunder Bay. Ontario 

B3 Variations in the Response of Fish Population 33 

Characteristics Lo Environmental Changes K. R. 

Munkittrick* and D. G Dixon. Department of Biology, 
Universityof Waterloo. Waterloo. Ontario. 

B4 MiExaminationof ChronicToxicity of Thiocyanate to 47 
Freshwater fish for the Development of a Water 
Quality Criterion D.G. Dixon, R.P. Lanno" andS.D. 
Kevan. University of Waterloo. Waterloo. Ontario. 

B5 Potential Role of Polycycl ic Aromatic Hvdrocarbona 55 
in the Development of Liver Tumors in Fish from 
PollutedSites of LakeOntario G M Kirbv. I.R. 
Smith, C.Thorr.. K.W Ferguson and M. A. Haves", 
Univeristyof Guelph, Cuelph Ontario. 

B6 Plant BloBssays for the Detection of Envlroonental 67 

.Mutagens InanAquatic Environment w.F Grant. 

Department of Biologj', York University, Downsview. 

B7 Effects of Temperature and Field Procedures oo PCS 79 
8 1 oaccumu 1 a 1 1 on InElliptio Complanata A Melkic* 

andV. Bollin, intergrated Explorations, Guelph. 

Abstract Page 


Oral Presentations 

D8 Biomonitoringt chemical Dependent Quantitative 67 

Relationships for the Body Burdens of Organic 
Chemicals in Aquatic Biomoni tors F Gobas", R 
RusseliandG, Haffner, Great Lakes Institute, 
Univerisity of Windsor, Windsor, Ontario. 

B9 Biomoni tor ing Protocols for Adult Aquatic Insects: ' ' ■ 
Seasonal Availability. Sample Size and Sens i tlvity 

2.E, KovatsandJ. J.H. Ciborowski-, Dept. of 
Biological Sciences. University of Windsor. Windsor, 

BIO An Ecosystem Approach to the Monitoring of PCB' s in 125 
Pristine Ontario Lakes CD- Metcalfe" andC-R. 
Macdonald, Trent University. Peterborough, Ontario. 

B11 Metal Contamination of Wetland Foodchains In the Bay 133 
of Quinte. Ontario A. Crowder", w Dushenko and J 
Greig, Dept. of Biology, Queen' s University. 
Kingston, Ontario. 

B12 An Over view of Aqua tic Environmental Research In 153 
Quebec M Slivitsky. INBS-E.\L", Ste. Foy. Quebec. 

B13 Development of an Improved System for the Application 155 
of Powdered Active ted Carbon in Water Treatment 
Plants H. Donison', A. Benedekand J. J. Bancsi. Zenon 
Environmental Inc. , Burlington, Ontario, 

B14 Municipal Utilization of Water Demand Management '" 
Strategies in Ontario Municipal i ties R. D 

Kreutzwiser* andR. B. Feagan, Dept. of Geography. 

University of Guelph, Ontario, 

Abstract Pag® 

Oral Presentations 

B15 A Preliminary Study toDetermltie the Feasibility of "I '9 
Medium Pressure Mercury Lamps for Disinfecting Low 
Quality Wastewaters G.E. Whitby andG. Sakamoto, 
Trojan Technologies Inc. , London. Ontario, andC. 
Palamateer", Envioronment Ontario. 

B16 Characterization of the Fecal Indicator Bacterial 247 
Flora of Sanitary Sewage with Application to 
Identifying the Presence of Sanitary Waste In Storm 
Sewers P.L. Seyfned". T.Bleier, V.XuandR. 
Harmandeyan. university of Torpnto, Toronto, 
Ontario I 

B17 Landsat-5TM Spectral Responses for Lakes Acroas 269 

NortheasternOntario JR. Pitblado, Geography 
Department. Laurentian University. Sudbury, 
Ontario. i 

' 9Sfi 

B18 Relationshlpof Mercury Levels In Sportflsh with Lake *•' 

Sediment andWater Quality Variables CD wren, 

B.A R. Environmental. Quelph. Ontario. 

819 Trend Analyais Procedures for Water Quail tyTloie 303 

Series A. I. McLeod*, andK.w, Hipel. McLeod-Hipel 
Associates Ltd. . London. Ontario andB. Bodo, 
Environment Ontario, 

B20 t'se of a Bromobenzoate for Cross -Adaptation of 31 1 
Anaerobic Bacteria In Lake Ontario Sediments for 
Blodegradatlonof Chlorinated Aroma tics M 
Urbanek". T. Strycek. R.c. wyndham and M. Goldner. 
University of Toronto, Toronto, Ontario. 

Abstract Page 

Poster Presentations 

BP1 The Effects of Agricultural Drainage on Sediment and 31 9 
Water Quality Loadings W. E. Watt. Department of 
Civil Engineering, Queen' a University. Kingston, 

BP2 WATQUAS 2.0: AnExpert System for the Water Quality 323 
Assessment of Ontario Rivers W.C. AllisonandT. E. 
Unny. Department of Civil Engineering. University of 
Waterloo, Waterloo, Ontario, andL. Logan, 
Environment Ontario. 

BP3 Geochemical Characterization. SizeFractlonatlon 325 
and Bioavailability of Trace .^letal Particulate 
Associations in the Don River L Warren and A. P. 
Zimmerman, Department of Zoology. University of 
Toronto, Toronto, Ontaj:io. 


BP4 The Investigation, E\'aluationandRecommendationof 

Biomoni tor ing Organisms for Procedures Development 
for Environmental .Monitoring C.A. Jefferson, Curry- 
Jefferson EInvironmental Services, Port Perry, 

BP5 The Ontario Inland Lakes Program and Management of 329 
Blue-Green Algae: Three Whole Lake Treatments In 1988 
H. VandermeulenandK. H. Nicholls, Water Resources 

Branch. Environment Ontario. 

BP6 Characterization of the Grazing Fauna Within Five 333 
Sof twater Lakes With Respect to Accumulations of 
Metaphytlc Filamentous Algae P.MStokes, E. T. 
Howell and R. L. France, Institute for Ernvironmental 
Studies. University of Toronto, Toronto. Ontario. 

Abstract Page 

Poster Presentations 

BP7 Sedimentary CKrysophycean Cygt Assemblages as ^^' 

Paleolndicators in Acid Sensitive Lakes M Rybak and 
I.Bybak, .\BECO Canada Inc . , Ottawa, Ontario, andK. 
Nicholls. Environraent Ontario. 

BP8 Factors Regulating Contaminant Levels In Forage Fish ^^' 
Species C.E. Herbert and G, D, Haffner, Great Lakes 
Institute. University ofWindsor, Windsor, Ontario, 

BP9 The Isotopic Composition of Upland Forest Soil 345 

Sulphate D.R. Vaji Stenipvoort and P, Fritz. 
Department of Earth Science, University of Waterloo. 
Waterloo, Ontario. 

BP1 Recent Trends and Historical Changes In Water Quali t>- 349 

of Lake >luskoka M. Rybak and I. Rybah. ARECOCanada 
Inc. , Ottawa. Ontario. andK. Nicholls. Environment 

BP11 In-Situ Determination of recal Indicator Bacterial 3^1 
Survival in Agri culturally- impacted Water sheds 

M.J Walters. Lake Simcoe Region Conservation 
Authority. Newmarket. Ontario. 

BP1 2 Development of an Acute and Chronic Sediment Bioassay 355 
Protocol Using Larval Mayf 1 ies and Juveni le Fathead 
.>Iinnow5 G. Kranizbergand R. Pope, University of 
Toronto, Toronto, Ontario, 

BP13Three Hour Pulse Exposure of Potass iumThiocyanate to *J0* 
Rainbow Trout Eggs Before and After Water Hardening 

S. KevanjIjidG, Dixon. University of Waterloo, 
Waterloo. Ontario 


5cienr:e-Bas«'i Ir.novatian 

Sci«nce-bas«d Innovation ia cci:ical In :od»y' a global economy to sustain 
and •rvhanca a nation's prosperity. In seaitiaa to s'jstaln and «nh»n=i 1-.3 
pco«p«rlty by participating In a groving vol'jffl* o£ vorld trad*, larga and 
snail econooiaa, faca critical pcoblems of adapting :heir institutions, 
pellcias and practices to a radically nav «nvironniani. Kay alamtnts ot 
this anvironaient ara that vorld trade now occurs In a global economy in 
which the interwtavlng of science. er-Bineeri.-.g and tachnology has acguirtd 
the povar to transform the ccsiparaciva advar.taga and prosperity of nations. 
With the scait, scope and openness of the Internacioral enierprisa of 
scianca, the tranaf arabtlicy- o£ tachnologies and the mobility o£ capital, 
science-basad innovation has becoEe a dcivinj force for the technological 
and corporate change that estates nev tradeabla goods and services- These 
conditions are radically diflarent from those of the laduairial RevolaClon. 

In a modern econosny the sector vhich produces tradaable goods and services 
supported by the firs: service sector at Einancial, legal, energy, 
transportation, cooaunicatlon systens, etc., generates the Ineoaie that 
enables a country to Invest In the second ser'/lce sector of education, 
health care and other parsonal and social benefits. (Figure 1). In some 
countries fina:icial institutions have been operating In a nar.ner that 
hampers the developnents in the tradeable goods and services sector. 

To participate in the global tccnomy driven by science-based innovation it 
Is essential that, on a national or regional basis, the pyramid of research 
capacity {In terms o£ knowledge flov) that leads to tradaable products and 
services has integrity, chat is, that there be a reasonable balance of 
capacity throughout the pyraal<l> (firara 2). 

Increasingly science-based innovation requires a strong long-terx applied 
research capacity, particularly in relation to anarging generic technolo- 
gies, that Is industry-based ajid controlled. This capacity has to be 
linked to a high quality fundamental research base and a strong market 
focused devtlopoent capability. 

L-arge and snail countries in different stages of development faces problems 

1) achieving structural integrity necessary for science-based 
Innovatiao suitable for their Halted resources of people and 
money, and 

il) using their United resources effectively. 

TrsdeabU Goods and Services 

• Liule direci job cruucn 

So^ct Sector I 

■ Finance 

. Communication! 
> Entrjc 

t Some aidetslt lervtcci 
aicr.ier sf world clua 


Service Sector n 

> Health 

I Education 

> Social S«f ^ ico 

> Leisure 

I Recreaiion 

• Dcpcr.(l*r,s on iacartw 
ftom ^A) 

• Sccter of job creatim 

• Low to ftijfiN iluUBdjobs 

• bf uctice* qiuBty of (A) 
and (3) 

FliTjre 1 : A Sinple y.cisl cf tha Economy 

In today's global economy It is impoctant to understand tha ralatlonshlp 
betveen Innovation In the production of tradaable goods and services and 
Che santracion of Income. A slaipla model in terms of stating the key 
issues Is given in Figure 1. This Bodel segsients the economy into three 
blocks labeled (A) Tcadeable Goods and Services, CB) Service Sector 1 and 
(C) Service Sactoc 11. The major source o£ Income which sustains our 
standard of living comes from sector (A) Tradeable Goods and Services. 
Canada'* current standard of living requires very substantial volumes of 
trade Into world Markets. In the globally compaci tivt Barket of today a 
nation must be concerned with maintaining and enhancing the environment it 
creates for business and Industry that can Innovate in Che production of 
tradeable goods and services. To function affectively, auch enterprises 
require a high quality service sector, nanely (3) Service Sec'oc I, 
coaprising such services as finance, legal, energy, construction, 
communications, transportation, distribution. It is the combination of 
this business service sector (vhlch produces some tradeable servlc-s) and 
Che sector directly producing tradeable goods and services which generate 
tha primary Income of a region. 

It is the income generated by the foregoing activities which allows the 
•xpansion of (C) Service Sector 11 that Is concerned vith personal and services. The social service sector includes health care, 
aducat.on, conifflu.-.ity services, leisure and recreational activities. Our 
capacity to sustain and Uprove the services and opportunities depe.".ds on 
th« capacity of sectors (A) and (S) to generate tha necessary income. 


The Sdence-Based Prramid of Research 


Compcditve or 

ShD«-ienB AppJiod 



Product 3t Service 

Fi^re 2 

Research as »n •Itment related to the ouerall process of Innovation, can be 
brolten down into three primary conponents that must be linked together to 
be effective: 


Basic or fu ndamental 

research vhich it uiually characterized by the 

primary objective being the generation of nev knovledge 
and understanding abou: nan end the vorld around us. This research is 
long-tem (usually on a tiaie-scale of 10 years or aote), and has a high 
level of uncertainty in ttrais of what the results vill be- In Vestern 
culture, basic research is primarily university-based and seldom 
results In knovladge that is of inmediate eoramerclal value. The 
Itnovledge gained from such researth is rapidly and videly distributed 
to scientists throughout the vorld through publication in scientific 
Journals. Because the results from this research are, or have been, 
considered a public good, this type of research has been financed 
primarily by the public sector and private benefactors. Increasingly, 
hovever, vhen knowledge contributed by basic research is critical for 
nev product development, industries tre becoming involved in basic 
research [OECD, 1987). 

2. *oDli< d res«arg:i = I.i •oat cauntries. «ppli«<i resmarth .s nalrly 
carritd ou-- HT industrial oi icv«rnm«nt laaocatcries, but ;n soma 
countrlaa. th€rt is aubs-.antlai -jnlv^rsitv Involvament vl:h rMpe-:; ':3 
longer term resMfth, p»r:ic'jl»rly in schools at ■ngineflrlng, msdisine 
and aanageaent. Applied :asMrcn ii»s a stratejlc :3rjac and attespts: 

10 «xtend lh« scop* of undecatandir.g af materials and 

to deceraint how tht accjinulatad 'ooviedgs Jrsia basic 
ras«atch, axt«nd whert necessary by foc-jsad spaclaiized 
resaacsh, can be used to develop a potential nev product 
or s«r/lces, or 

to dsteraine hov to modlEy and Upcova th* perfiJrT.anc« 
of axlstlng products or services to sustain their 

Applied research vhich is inedlu.-n to lonj-term (on a tine scala of three 
to ten years), also has a significant level of uncertainty, but because 
It is targeted, chare is » probability chat there vill be econo'sic 
benefit. The means for the financing of this research vary. In some 
sectors such as tha phacnaceutical and chemical fields, the research Is 
largely funded by the private se-tor primarily through the benefits 
froa patent protection, vhereas in fields such as aircraft and 
•lectronics, there has been a mixture of public and private financing. 
In some cases a monopoly position (e.g. AT&T and Bell Laboratories) has 
encouraged the funding of longer-terx applied research, but there are 
fev examples of the private sector being able to finance longer tern 
applied research vholly from its ovn resources unless there Is 
flffactlva patent protection or the busi.iess has a monopoly position. 

It is common in son* sectors to associate tha processes of engineering 
design and development of a product or service as discussed above, vith 
the term development or developmantil research. 

3. Developmental research is research thati 

- use of the fruits of applied research specifically to 
create a nev marlcetable product or service, or 

• improves, through a series of small steps of innovation based on 
■tate-of-the-art luiovledge, an already existing product or 
service, or 

- enhances th* ease of production of a product or the provision of 
a service. 

This type of research has the least uncertainty, is carried out on a 
time scale of less than three years, and has the highest pcobaiilUty of 
economic benefit, developmental research is mainly financed by the 
private sector, although there are exceptions in which there has been 
substantial public financing. 

Th« foragoing ca:egoi-i« of resair:h can b« c«or«3«n:ed by cht pyramids 
ahovn in Fifjra 2. At th« narrav p««x o£ aach pyraaid la a pcorfuc: or 
3er-/Ic«, a spaclflc artifact oi technology designed to perform a particular 
function in a »arlta;. ?rom Its p««i(, each pyramid expands througli tti rhr«« 
prliTiacy categorias of rastarch to a broad basa in basic rastarch. Thi 
category "applied research* has been segnentad into cvo slicss labeled 
coBpetitiva Cshort-tam apolied) ar.d pc«-;oii?eti:i-'e (long-tera applied). 
Competitive applied rasaar'-h is that vhlch has direct pcoprietar-/ valua to 
the business. Pra-coapetitive applied research is :ha: vliich is sar.erally 
usafui in sectors of industry (This reaaacch is octan concerned vhit 
can ba called generic tachnology). The relative vldth of aach 3li=« across 
the pyramid suggests the range of generality oi the luiovladge associated 
vith It. The overlapping of pacts of tha pyramids indicates that as one 
reaches tovards the acientiiic roots pertinent to the development of ft 
particular product, the Itnovladge base becomes ralavant to a range o£ 
products. Indeed, the essence of basic science is that it seeks tor general 
principles of understanding vithin particular circunstances of study, 
vhareas engineering, through the technology it creates, sealu to realise a 
particular operational function in a marltet vithin the domain ot 
possibilities bounded by science. 

Science-based innovation then is innovation In vhleh the realization of an 
effective and competitive product or service utillias, through the focusing 
processes of the pyramid of research, tha full range of scientific and 

engineering understanding pertinent to the function o£ tha product or 
service in the marketplace. 

The classification of levels of research in tha rejearch pyramid of Figure 2 
Is based on the diverse literature on Innovation. Its pertinence tor older, 
large-scale, science-based industries is clear. Sovever, a key point today 
is that the research pyramid is relevant to all industry participating In 
the global economy of science-based innovation. 

SCIENCE, ENCINEERIMG AND TSCHNOLOGY" may be obtained by filling out the 
attached form. 



Stuart L. Smith, M.D. 


RockCliffs Research and Technology Inc. 

Novambec 1988 

As difficult as research can be, it is still more 
difficult to apply it swiftly for economic or social benefit. 
In addition to the usual obstacles to technology transfer, 
environmental research faces additional ones of a political 
nature. It behooves us to know a great deal more about how 
research is transformed into practical benefits and how 
environmental research in particular can be more rapidly 
applied. The improvement of the environment is an area 
where, with appropriate policies, economic and social 
benefits occur simultaneously. 

In supporting research activities, we cannot take 
for granted that application will naturally follow any 
improvement in knowledge. More attention needs to be paid to 
the incentive structures of research organizations, the 
relationship to our industrial sector, and the interaction 
with political decision-making. By acting now in some 
specific areas, we can help guarantee that today's researcli 
will produce timely and tangible results. 


Oral Presentations 


•F- ';W 



... - -■■■■ , . - - L-^ 



Carl Schenk 

Wator Resources Branch 

EnvironmenL Ontario 

Research needs related to Mater and the aquacic environment 
cover a broad range o£ concerns and issues. The needs identified 
nereafter reflect the Ministry's interests in establishing 
Improved treacmenc processes for the Province's drinking water 
liUpplles and the removal or neutralization of municipal, 
industrial and diffuse source wastes, developing a better 
'jnderstanding of cause-effect relationships in aquatic systems so 
that effort can be directed to deal more eftecLively with water 
quality impacts, and determining improved environmental evaluation 

For convenience, water research requirements have been 
yrouped into the following eight categories, involving 33 issues 
and about 130 specific needs; 

1) Industrial wastewater Treatment 

2) Municipal Wastewater Treatment 

31 Managing Non-Point Sources of Pollution 

4) Contaminant Fate and Transport Processes in Aquatic 

5) Impacts of Pollutant Discharges on Aquatic Systems 

6) DrinJtlng Water 

7) Effects of Acidic Deposition and Long Range Transport of 

a) Other 

Research issues have been altered and the specific needs 
adjusted for fiscal year 19B9-90 based on input from head office 
and regional staff throughout the Ministry. New issues include 
WAIO to investigate effects of intensive crop production practices 
on groundwater quality. WA14 to identify remedial measures to 
minimize the impact of lakeshore development, WA15 dealing with 
the aquatic effects of timber management practices. WA18 involving 
the development of models to address contaminant fate and 
transport in the Great Lakes, WA2! to model the impacts of 
contaminant discharges on aquatic biota, WA30 on the distribution, 
behaviour and effects of low level trace metals in aquatic 
systems, WA31 concerning the effects of organic contaminants 
associated with long range transport and finally, WA33 dealing 
with the spatial anayses of water quality. 

Clearly, the Ministry's top priority in water research 
relates to evaluations of the significance of hazardous 
contaminants and minimization of these contaminants as a threat to 
our natural waters and drinking water supplies. This emphasis is 
reflected by the vast majority of the 33 research Issues and 
related needs that have been identified. Studies carried out by 
universities and private sector consultants within the Ministry's 





AFRIL 26-26, 1908 

K.E. Day'. E.D. Onglay', R.P. Scroggina^ and H. Elsenhauar^ 

^Env Irannent Canada, Riven Rasearch Branch. 

National Water Research Institute. BurllnBton, Ontario 

^Environment Canada, Place Vlocant Kaasey, Hull, Quebec 

The nev Canadian Envlronnantal Protection Act (CEFA) togethar with 

other legislative initiat ivea. e.g.. Ontario's Municipal Industrial 
Strategy for Abatement program (MISA) , provides the policy framework 
to expand the traditional chemical approach for hazard asseasment of 
pollutants In Canadian waters and sediments to a combination of chemi- 
cal and biological toxicity tests for protect Ion of the aquatic 
environment , The benefits of incorporating biological testa and 

standarda into regulations are well recognized; however, there are 
reservations about the current level of acientlfic knowledge and 
practical expert ise aval labia to formulate and Implement such regula- 
tions at this time. 

Concerns have been ajtpresaed about the types of biological taata to be 
used, shifts in regulatory criteria due to Improvements In the 
knowledge-base, the role of governmanta in RiD relative to the private 
sector and technology transfer from govarnnwnt laboratories to the 
private sector, university RSD and conaultlng, etc. This concarn led 
to a national workshop, hosted by Environment Canada and chaired by 
the National Water Research Institute on April 26-28. 1989, at the 
Hattawasaga Inn, AlHston, Ontario. Called "Biology In the New 

Regulatory Framework for Aquatic Protection", participation included 
represent at ives from senior and technical levele of several federal 
government departments, provincial oEfteials, university faculty, 
regulated Industries, private consultants and laboratories, and muni- 
cipal wastewater treatment operators. The workshop was designed to 
promote discussion amongst key decision makers and experts from the 
various sectors on the imp lie at Ions, problems and economic opportuni- 
ties associated with Incorporating biology Into aquatic environmental 
regulations. The objective of the workshop was to provide advice to 
enable regulators to deal more affectively with the policy Implica- 
tions of biological assessment as a regulatory tool. 

Invited speakers provided an overview of biological toxicity tests in 
Canada as they relate to CEPA, MISA and the Post Control Products 
Act. Industry and the private consulting sector were Invited to 
respond with discussions of the benefits, problems and opportunities 
presented by these legislations. In working groups, the participants 
formulated a response to the following questions; what are the bene- 
fits and limitations of biological toxicity tests as compared to other 
possible methods and strategies, i.e., the chemical-specific approach] 
what is the role of the government in research and development of 
biological toxicity tests; what is the role of the government in 
monitoring and assessment? 

Full proceedings of the Alliston Workshop will be issued In a separata 
document by Environment Canada in 1989. 


Th« najor reeomniendst Ions srisiriB from the Workshop can be sijinni«rized 
•a to] lows: 

1. Governments must develop a proBram framework and outline policy 
statenenta required to set biology'baaed regulatory standards and 
to expedite th« development of standardized protocols for 
biological toxicity tests. 

2. Biological testing and monitoring must be integrated vtth chemiS' 
try In a mutt i-dlscipllnary manoar wh«D applied In hazard SBtssa- 
ment and regulatory control. 

3. SubJethal and chronic teats need to be developed to provide more 
sensitive monitoring tools for detecting toxic effects In ambient 
waters , 

4. Mechanisms for improved conmunicat ion, awareness and understand- 
ing of the applied uses of biological toxicity tests amongst the 
scientific community, regulatory agencies, industry and the 
public sector must be pursued. 

5. Government must maintain a strong research and development 
capability and continue to transfer technology developed in the 
Held of biological testing and monitoring Co the private sector. 

6. Government should provide guidance and show leadership in the 
area of biological testing and monitoring QjV.'QC, e.g., policies, 
guidelines, laboratory accreditation, toxicologist certification, 

7. Government is responsible for the maintenance of long-term 
ambient monitoring schemes while industry is responsible for the 
assessment of immediate areas of impact, e.g. , end-o£-pipe and 
mixing zone, with occasional auditing by the government. 



Basic Relationships and Simple Kinetic Modelling. 

L.S. Mccarty . ■* : G.W. Ozburn*; A.D. Smith", and D.G. Dixon- 

* Aquatic Toxicity Research Group, Lakehead University, Thunder Bay, 
■ Biology Department, University of Waterloo, Waterloo 


Some of the assumptions of commonly employed aquatic bloassays where 
toxicity or bioconcentration are estimated are reviewed. For many 
organic chemicals the link between bioconcentration-derived and 
toxicity-derived kinetics information, as well as the hypothesis that 
typically employed biological endpoints occur at relatively constant 
body burdens, can be exploited by means of a one-compartment, first- 
order kinetics model. when verified for the mode of toxicity and the 
general character of the test species, the model can be used to 
explore situations not explicitly examined in the original data. Such 
deterministic models can also be used, in the standard scientific 
approach of hypothesis formulation and testing, to formulate 
hypotheses to direct future experimental designs. Examples of 
applications dealing with mixtures of toxicants and intermittent 
exposure regimes are discussed. 


Large amounts of data concerning bioconcentration and toxicity have 
been collected for a variety of purposes including: monitoring, 
regulation development and/or support, legal requirements, and basic 
investigative science. Numerous chemicals, organisms and 
circumstances have been examined. Despite this the data available are 
often only of very restricted utility in new situations as the 
organisma or conditions or both are substantially different. 

This is in large part due to the fact that aquatic toxicology has 
evolved largely a descriptive discipline. Studies examine the problem 
primarily by reporting a detailed description of the circumstances of 
the test and of its outcome. This stochastic approach attempts to 
correlate changes in results with changes in one or more experimental 
parameters without necessarily attempting to explain an underlying 
mechanism. Such descriptive methods are ultimately handicapped by the 

lack or an underlying general theory. 

To be able to further exploit the data and increase the understanding 
of the phenomena aquatic toxicity studies must be directed by the 
traditional scientific approach: hypothesis formulation and subsequent 
testing. Deterministic models must be employed so that changes in 
results are correlated with varying experimental parameters, as in the 
case of the stochastic model, but also changes are also examined in 
terms of at least soae of the fundamental biological and physical- 
chemical processes involved. 

In this paper some of these basic concepts and assumptions will be 
reviewed, a examination of how simple deterministic models may be 
constructed from currently existing toxicity and bioconcentration data 
will be carried out, and hypothesis formulation and testing will be 
briefly explored. 

Four general areas need to be examined: toxicity tests, 
bioconcentration (BCF) tests, toxicokinetic modelling, and the 
constant body toxicant concentration concept. 

The primary objective of the toxicologist carrying out the toxicity 
bioassay is to determine the potency of the toxicant being examined 
relative to the potency of those which are already known (Bliss, 1957; 

Filov et aj.. . 1973) . There appears to be two basic explicit 
aasumpt iona in this essentially stochastic approach: 

1. a dose-response relationship exits. I.e. that the biological 
response of the organism exposed to the toxicant is some function 
of the amount of toxicant to which it has been exposed, and 

2. the distribution of tolerance of the measured biological 
response of test organisms to the toxicant is known. 

The assumptions require some further explanation. The dose-response 
relationship is the fundamental assumption of toxicology but to employ 

it some additional assumptions are implicit. Some measure of the 
amount of toxicant to which the organism is exposed, i.e. toxicant 
external to the organism, is being used as a surrogate for the amount 
of toxicant In the organism at the aite(s) of toxic action which is 
the ultimate cause of the biological response. 

Due to the complexity of non-equilibrium, non-steady-state kinetics it 
is usually assumed, implicitly, that a steady-state equilibrium 
occurs, or at least could occur, between the toxicant external to the 
test organisms and that which is in the body of the organism. Thus at 
"threshold" or "incipient" toxicity levels, with either lethal or non- 
lethal biological responses, it is valid to compare the potency of 
different toxicants (Sprague, 1969, 1970; Filov et al. . 1973). 

Also implicit is that the metabolism or biotransformation of the 
different toxicants which are being compared is similar, and usually, 

The assumption of a normal distribution of the logarithm of the 
tolerance {log-normal) in the exposed population is commonly made as 

is facilitates statistical analysis (Finney, 1978). 

Bioconcentration tests do not, as mentioned earlier, focus on the 
response aspect of the dose-response relationship, but rather 
emphasize the dose component. In fact, if a slightly enhanced view of 
the dose-response relationship - exposure, kinetic, and dynamic phases 
- is employed (Ariens, 1980) it is clear that most bioconcentration 
research is directed to the kinetics aspect - uptake, distribution, 
biotransformation, and elimination - of the relationship. Thus 
bioconcentration tests are inherently based on a dynamic model 
approach . 

A variety of models of various levels of sophistication can be 
employed (Dedrick, 1986), but the common approach has been to use the 
simplest: the one-compartment, first-order kinetics model (ICFOK). 
Despite the simplicity the model has been used and discussed for a 


variety of chemicals and aquatic organisms ( Mancini, 1983; Spacie and 
Hamelink, 1983; Hawker and Connell, 1985; Barber fii al*, 1988). 
Figure 1 sunuDarizee the nathenatical description of this model. 

Figure l. 

One-Compartment, First-Order Kinetics Model; 
Mathematical and Graphical Form 





Wloss Ct| 

6 . 




">. \ 

A . 



^ \ 



\: >■ r 


■kr?M ^ 
Cw«KI/lc2*[i'e 1 





— i 

1 , ^ ) — 

— f -H 

■i. 5 6 


It is becomes clear that the design and interpretation of aquatic 
toxicity tests is rooted in a stochastic model of the phenomena, while 
the basis of the bioconcentration test is a deterministic model , In 
previously published work (McCarty, 1987a, b) it has been suggested 
that, at least for poorly metabolized, neutral, narcotic organic 
chemicals, information from both toxicity tests and bioconcentration 
tests could be integrated since, essentially, each was focused on an 
opposite side of the same coin. That is to say, the kinetics should 
be similar in both cases and differences that occur should primarily 
be a result of the difference of the endpoints being employed. Since 
the biological response ie a function of the amount of toxicant which 
enters the body of the organism both bioassay endpoints will be 
referenced to a common basis, the whole body burden of toxicant. 


The whole-body concentration or burden - an internal level - is an 
approximation of the toxic dose which is more closely associated with 
the biological response than the concentration or dose of toxicant 
which is applied externally to the test organisms. The toxicant bound 
to the receptors at the site of toxic action is a much more accurate 
measure, although considerably more difficult to estimate. Thus the 
body burden or concentration, incorporating adjustments for known 
influencing factors such as varying 1 ipid levels, will be used as a 
reasonable first approximation estinata of the actual effective 
internal dose. 

Concentration, either internal or external, is itself is an 
approximation for the effective dose in the organism since, as 
elucidated by Ferguson (1939) , it is the activity of the number of 
molecules of toxicant in the organism and not their actual number 
which is thought to be most closely related to the biological 

Therefore, in toxicity bioassays, within a common mode of toxic action 
and species and condition of test organism, the body burden of 
toxicant associated with the toxic endpoint being employed should be 
relatively constant (Connolly, 1985) . Support for this comes from 
QSAR work, McCarty (1986, 1987a), working with toxicity and 
bioconcentration data, reported that for the acute toxicity of some 
neutral, narcotic, chlorinated hydrocarbons the body burden associated 
with 96 h LC50 estimates appeared to be about 2 mmole/L internal 
concentration for fish of about 5% lipid content (2 mmole/kg if fish 
density is 1.0). 

More recently van Hoogen and Opperhuizen (I9S8) actually measured body 
burdens of chlorobenzenes at the 96 h LCSO and found a body burden 
estimate of about 2.5 mmole/L in fish with a SI lipid level, which is 
in remarkably good agreement with the predicted value. 

It must be noted that, in the typical toxicity test, the toxicity body 
burden is being measured indirectly, by the mortality response of the 


exposed population, and this and other variability associated with the 
estimation of tolerance distribution by this means will be 
incorporated body burdens estimated in this way. 

For the bioconcentration bioassay the body burden is actually measured 
in the test so estimates should be more accurate. For many commonly 
studied organic chemicals it appears that the ratio of the exposure 
concentration to the body burden is relatively constant over a range 
of exposure concentrations and follows the well-established 
relationship of Increasing body burden at steady-state equilibriim as 
a function of the lipophilicity of the chemical (Mackay, 1982). 

It becomes clear that the examination of the relationships between 
data derived from standard toxicity and bioconcentration bioassays 
requires accepting several assumptions. Rather than detail these 
assumptions further we will simply state the assumptions that are the 
basis for the working hypothesis. 

1. The uptake and elimination of many organic chemicals, especially 
neutral narcotics, can be reasonably approximated by a one- 
compartment, first-order kinetics model which is independent of the 
biological activity or response endpoint under examination, 

2. for the case of neutral, narcotic organics with the same mode 
of toxic action the biological response endpoints estimated by the 
statistical response of the exposed population, and commonly 
employed in aquatic bioassays: lethality (eg. threshold LC50) , 
growth, and inhibition of reproduction, are a function of a 
relatively constant body burden of the toxicant. 

As well constant bioavailability of the toxicant and no uptake of 
toxicant from the diet, negligible biodegradation and growth for the 
duration of exposure is assumed. 


■,■*•--■. •'^V/ 

•■yw'-ivV-*-. -" 


Toxicity information for 1,4-dichlorobenzone were obtained in 
continuous- flow, acute toxicity tests with juvenile American flagfish 
(Jordan pll^ florldae Goods and Bean) of an age of 2 to 4 months (Smith 
et al. ■ manuscript). The estimated 96 h LC50 was 2.05 mg/L (14.0 
ymole/L) . Samples from a series of tests indicated fish sizes ranged 
from 0.3 to 5 g (typically 0.5 to 4.0 g) and lipid contents varied 
from 7 to 16 per cent (typically 8 to 12 %) . A non-toxic (less than 2 
% of the LC50) level of acetone was used as a solublllzlng agent. 

The time-toxiclty data were used to obtain estimates of K2, the 
elimination rate, and the factor l/Cf*Kl/K2, which is the product of 
the inverse of the fish toxicant concentration times the 
bioconcentration as discussed by McCarty (1987b). The nonlinear 
routine from the PC-based statistics program SYSTAT 4.0 was employed. 
Figure 2 sunuoarizes the approach and the results. 

Figure 2. Mon-Linear Curve Fit of Acute Toxicity of 
l,4~Dichlorobenzene at Various Times 


mMoie/L) American Fiagf'sri 

« * *■ 





^1/Cw = l/Cf*kl/k2 (1-e ) 

Cw : IC5Q waier level 
I : lime ol LC50 estimale 



Kl/k3 -. BCF 

1:2 estimate ■ tJSi pet Hour 



l/Ct'fcl/fcS esiimale : 59 
-H \ 1 1 1 

iE (days) 


ThB bioconcentratlon data were obtained with fish of Binilar 
characteristics employed in the toxicity testing. The fish were 
exposed for 28 days to a measured concentration of 2.7 nq/1. (0.018 
(inole/L) of 1,4-DCB in 79 mg/L acetone followed by a 14 day depuration 
period. (ATRG, 1987). The exposure water concentrations and body 
burden estimates at different times were analyzed with the BIOPAC 
program to generate kinetics constants and a bloconcentration factor 
based on the whole body toxicant level. The BIOFAC program is 
essentially a nonlinear curve-fitting routine customized for use in 
analysis of bioconcentration data {Blau and Agin, 1978). The data and 
results are presented in Figure 3 . 

Figure i. 

1,4-Dichlorobenzene Bioconcentration Kinetics: 
BIOFAC Model Analysis 

BODY BURDEN (umole/g) 

1 t)E-D3 




^ J ^ ■ -fl 

^ a ■ ^ 

Flaghsn approximately 0.5 g, 

9K lipid Vb 

ki '- ?9t oay''* 


lc2 =0 98 oay"^ 

BCF '- 295 


Cw = 0.Q1B umoie/L 
1 1 \ — _j 

— h 1 1 





TIME (days) 

Hypothesis Formulation 

since our first assumption is that kinetics are independent of the 
biological response being investigated it follows that, allowing for 
variance due to experimental factors and organism sensitivity, the 


kinetics fron a bioconcantration test should be very sinilar to thosa 
obtained in a toxicity test. Thus we have taken the kinetics 
information from Figure 3 and nodeled the time course of toxicant 
concentrations in the body which should occur when flagfish are 
exposed to varioue water levels of 1,4-DCB as indicated in Figure 4. 
Also indicated is the approxinate body burden associated with the 96 h 
LC50, 2 nunole/L. 

rlgura 4. 

Hypothesis; Uptake of 1,4-Olchlorobenzene at 
Various Hater Concentrations 

Bofly Suraen (mmoies/L 

100 t5C 

Time inours 

Test of the Hypothesis 

In Figure 4 the tine-course of body burden accumulation has been 
modeled for several endpoints; however, the most practical endpoint 
for use in testing the hypothesis is the threshold LC50 endpoint as it 
is currently tha aoat well-defined . Although the body-burden 
estimate associated with this endpoint would more realistically 
represented by a confidence interval about the mean estimate, further 
refinement is currently being carried out and this value is all that 


is currantly available. 

Figure 5 is the one-compartiaent, first-order kinetics nodel output for 
a 96 h exposure to a concentration of 0.014 mmole/L of 1,4- 
dichlorobenzene, the estimated 96 h LC50 for these organisms. The 
model was run under three sets of parameters, identified as BCF, 
TOX-0, and TOX-E, and the results are presented in the figure. 

The model output labelled BCF is based on information - kl, k2, and 
bioconcentration factor (BCF) - taken entirely from the 
bioconcentratlon bioassay data. 

rlTuze 5. 

Hypothesis Test: Uptake of 1,4-Dichloroben2ene 
at Estimated LC50 Water Concentration 

Body BurOen - mmote/L 

iCFOK Mooei oulpui 1,4-aicriiofooenzene 
Fiagdsn; 0,5 lo 4,0 g, fl-lJK iipiO 


2* 36 48 60 17 9* 96 lOB '?0 132 144 

TIME - nours 

In the observed bioconcentration-toxicity (TOX-O) output the 
elimination constant, k2 , was derived from the non-linear curve 
fitting of the time-acute toxicity data presented in Figure 2 while 
the BCF value is from the bioconcentration results. The uptake 
constant kl is derived from these two values. 


For the case of the estimated toxicity (TOX-E) output the k2 estimate 
is from the toxicity curve-fitting data while the BCF parameter 
estimate is calculated from Mackay's (1982) relationship for 
bioconcentration and Kow. A log Kow estimate tor 1, 4-dichlorobenzene 
of 3.52, a measured estimate from the MED CHGH 3.53 database (Leo, 
1988), was used in this calculation. The constant in the Kackay's 
equation was changed from 0.05 to 0.10 to reflect the higher lipid 
content of the Clagfish used in this study versus. Fish lipid levels 
were likely closer to 5 % in the data used by Mackay {19B2) while, as 
noted earlier, 10 * is a more reasonable approximation for the 
flagfish used in this study. 

From inspection of the figure it can be seen that all three of the 
curves plotted are in very close proximity to each other. In addition 
the estimated toxicant concentration in the fish are close and compare 
very favourably with the estimates of toxicant body burden estimated 
for acute toxicity in other studies - 2 and 2.5 mmole/L - noted 


Although this study is not an exhaustive evaluation of the problem it 
would seem reasonable, given the close similarity of the different 
uptake curves and the agreement of the model -estimated toxic body 
burdens with an independent estimate of body burden for the same toxic 
endpoint, to conclude that the hypothesis which was proposed has been 
validated for the circumstances considered. Thus it appears that: 

1. Kinetics information from toxicity and bioconcentration tests 
is very similar, and 

2. a one -compartment, first-order kinetics model appears to be a 
the deterministic model which, at least in the first approximation, 
is adequate for the study of toxicant kinetics and dynamics in 
aquatic bloassays measuring both acute toxicity and 
bioconcentration . 


These conclusions are limited to the circumstances which were 
examined; however, since the deteministic model approach has b«en 
validated it can be employed as the basis for the investigative 
procedure of hypothesis formulation and testing. 

Two important areas can be addressed in this way. First, currently 
available bioassay results can be better interpreted and interpolated 
while results for new chemicals and/or circumstances can be predicted. 
Second, the toxicological significance of body burdens or tine series 
of changing body burdens of organisms collected in the field can be 
estimated and compared with data obtained under more rigorously 
controlled situations. 

An example of an application of the hypothesis formulation and testing 
procedure applied in the first area discussed above appears in Figure 
6. The kl and k2 values for 1 , 2,4-trichlorobenzene and 1,2,4,5- 
tetrachlorobenzene were obtained from bioconcentration tests carried 
out exactly as described earlier for 1,4-dichlorobenzene, which is 
also included in this figure. The model output indicates hypothesized 
body toxicant changes at a exposure concentrations ten times that 
employed in the original bioconcentration bioassays. In addition the 
sun of the body burdens of the three chlorobenzenes is obtained at 
each san^ling tine and plotted as the total chlorobenzene body burden. 

Experiments could be designed and carried out to test several aspects 
of this hypothesis. Bioconcentration bioassays could be carried out 
at water concentrations ten times the original water concentrations to 
examine success of the model predictions for the time course and 
ultimate body burden for each of the three chlorobenzenes. As well a 
study of the bioaccumulation of a mixture of the three chlorobenzenes 
would detemina the success of the model in prediction the 
bioaccumulation of a mixture. 

In addition, since body burden levels at the higher exposure levels 
used in the model output are of the order of nagnitude of that 
associated with substantial acute lethality, toxicity tests could be 


riTur* C. Hodal Output for a Mixture of 3 Chlorobanzenea 
Body aurd«n - mtiitmoieU 5 lo 4.0 g, 8 - \2% lipid 




Cw Cf 

12 1 


0.01B 0.54 




0-021 4 10 

^^ — ^6 7 9 


0014 5 18 









(CI '- 9 B) 
, 1,2,4, 5-TeCB 

A - 

/ ^^^ 

— — — \\^ 



2 - 








k 4 — , -( 

' 1 1 ; — 

1 . ■■ 1 M 

96 120 '*^ 1GB 192 

Time - hours 

2\h 240 3E4 see 

carried out, both with single chemicals and with a mixture, to 
determine the relative toxicities. 

If the model reasonably describes the circumstances a difference in 
the mortality caused by various chlorobenzenes should be apparent. 
For example, population nortality of say 20% for 1,4-DCB and 50% for 
either 1,2,4-TCB or 1,2,4,5-TeCB (these latter two are likely so close 
as be distinguishable under the test conditions) might be expected, or 
at least differences in mortalities In this order. If the toxicity of 
the three CBs are simply additive then a toxicity test employing a 
mixture of the three would be expected to produce a higher percent 
mortality, say 80%, than the single chemical exposures and the onset 
of mortality might be expected to be earlier than the others, as 
suggested by the shape of the Total CB output curve in Figure 6. 

Another aspect which could be investigated is the situation where the 
exposure to the toxicant is not continuous i.e. intermittent or pulse 
exposures (Holdway and Dixon, 1985). Since a kinetics-based dynamic 


nodel is being employed it is a relatively simple mathematical 
exercise to predict the body burden through various changing exposure 
levels. The test of the hypothesis is an actual experiment to see if 
the biological response reflects the responses expected and/or if the 
body burdens follow the predicted course. Figure 7 illustrates the 
hypothesized body burdens for a situation similar to that shown in 
Figure S but with the exposure being of an intermittent or pulse 

Figure 7 . 

Hodel Output: A Mixture of 3 Chlorobenzenes 
with a Intermittent Exposure Segina 

BOfly Bufden - mmole/L 

Fiagiisn o 5-4 g, B-12« lipid 


6 - 

Tolal Cniorobenzenes 
1,2,4, 5-TeCB DOOH mmoie/L 
-1,3,3-TCB 0.0021 mmoie/L 
1,4-DCB' 0014 mmoie/L V 

lOOx original 8CF aata 
t6 nouf miervais^-. 


— * 1 1 — "^ 

18 n 96 i!0 

TIME - hours 


nature, 16 hours of exposure to toxicant followed by 16 houra of 
exposure to clean water for a total of 6 cycles. Experiments could be 
designed to examine body burdens and toxicity of the 3 chlorobenzenes, 
both individually and in mixtures, in a manner similar to the 
continuous exposure investigations. 

Experimental data examining individual chemical toxicity, mixture 
toxicity and bioconcentration, and intermittent exposure toxicity is 
currently being collected and/or analyzed In our laboratories and we 


hope to report on these actlvitlea in the near future. 

We trust that, within the limitations specified, we have demonstrated 
how toxicity and bioconcentration bioassay data are related and how 
this infomation night be exploited within the confines of a 
deterministic model based on the concepts of one-compartnient, first- 
order kinetics and the association of a biological response with a 
relatively fixed body toxicant burden. Furthermore, we trust that we 
have illustrated how this approach can be used to in hypothesis 
formulation and testing, both in interpolating and extrapolating 
existing data as well as in experimental design. 

Acknowledgements : 

The authors would like to gratefully acknowledge the financial support 

of the Ontario Ministry of the Environment. 


Ariens. E.J., 1980. Design of safer chemicals. IH E.J. Ariens (ed.). 
Drug Design, Vol. 9. Academic Press. New York NY. 235 p. 

ATRG, 1987. Aquatic Toxicity Studies of Multiple Organic Compounds. 
Part 1. Chlorinated Benzenes and Chlorinated Phenols: Summary Report 
(Interim). A Report to the Ontario Ministry of the Environment and 
Environment Canada. Aquatic Toxicity Research Group, Lakehead 
University, Thunder Bay, Ontario. 277 p. 

Barber, M.C. , L.A. Suarez, and R.R. Lass iter, 1988. Modeling 
bioconcentration of nonpolar organic pollutants by fish. Environmental 
Toxicology and Chemistry 7:545-558. 

Blau, G.E. and G.L. Agin, 1978. A users manual for BIOFAC: A computer 
program for characterizing the rates of uptake and clearance of 
chemicals in aquatic organisms. Dow Chemical Co. Midland HI. 10 p. 

Bliss, C.I., 1957. Some principles of bioassay. American Scientist 


Connolly, J. P., 1985, Predicting single-species toxicity in natural 
water systems. Environ. Toxicol. Chem. 4:573-582. 

Dedrick, R.L., 1986. Interspecies scaling of regional drug delivery. 
Journal of Pharmaceutical Sciences 75:1047-1052. 

Ferguson, J., 1939. The use of chemical potentials as indices of 
toxicity. Proc. Roy. Soc. B 127:387-404. 

Filov, V. , A. Golubev, E. Liublina, and N. Tolokontsev, 1979. 
Quantitative Toxicology: Selected Topics. John Wiley and Sons 
New York, NY. 462 p. 

Finney, O.J., 1987, Statistical Hethod in Biological Assay, 3rd 
edition, c. Griffin and Co. Ltd. London, England. 508 p. 

Hawker, D.W. and D.W. Connell, 1985. Relationships between partition 
coefficients, uptake rate constant, and the time to eguilibriun for 
bioaccumulation. Chemosphere 14; 1205-1219. 

Holdway, D.A. and D.G. Dixon, 1985. Acute toxicity of pulse-dosed 
methoxychlor to juvenile American Flagfish, Jordanella f loridae Goode 
and Bean) as modified by age and food availability. Aquatic Tox. 

Leo, A. and D. Weininger, 1988. MED CHEM Database, Version 3,53. 
Pomona Medicinal Chemistry Project, Pomona College, Claremont, CA. 
Information provided by Dr. s. Broderius, U.S. EPA-Duluth. 

Mackay, D. , 1982. Correlation of bioconcentration factors. Environ. 
Sci. Technol. 16:274-278. 

Mancini, J.L., 1983. A method for calculating the effects, on 
aquatic organisms, of time varying concentrations. Water Res. 

McCarty, L.S., 1986. The relationship between aquatic toxicity QSARs 
and bioconcentration for some organic chemicals. Environ, Toxicol. 

Chem. 5:1071-1080. 

McCarty, L.S., I9e7a. Relationship between toxicity and 
bioconcentration for some organic chemicals. I. Examination of the 
relationship. IN K.L.E. Kaiser (ed.). QSAR in Environmental 
Toxicology - II. D. Reidel publishing Company, Dordrecht, Holland. 
Pg 207-220. 

McCarty, L,s., 1987b, Relationship between toxicity and 
bioconcentration for some organic chemicals, II. Application of the 
relationship. IN K.L.E. Kaiser (ed.). QSAR in Environmental 
Toxicology - II. D. Reidel publishing Company, Dordrecht, Holland 
Pg 221-230. 


Smith, .A. D. , A. Bharath, C. Mallard, D. Orr, J. A. Sutton, J. 
Vukmanich, and G.W. Ozburn, nanuscrlpt. The acute toxicity of 10 
chlorinated organic compounds to the American Flagfieh Jprtj a n gt l a 
floridae (Goode and Bean). Submitted to Aquatic Toxicology. 

Spacie, A., and J. Hamelin)c, 1982. Alternative models for describing 
the bioconcentration of organics in fish. Environ. Tox. and Chem. 

Sprague, J.B., 1969. Measurement of pollutant toxicity to fieh I. 
Bloassay methods for acute toxicity. Water Research 3:793-821. 

Sprague, J., 1970. Review Paper: Measurement of pollutant toxicity to 
fish II. Utilizing and applying bloassay results. Hater Res. 4:3-32. 

Van Hoogen, G. and A. Opperhuizen, 1988. Toxicokinetics of 
chlorobenzenes in fish. Environ. Tax. chem. 7:213-219. 



Effects of BiKcd aetal Bintng «aKte on whitR KUcker populat lonN.' i1evcto[«ent of 
a froaework to describe fish population responEe»> to nnviroiwufntdl change. 

K.R. Munkittrick' and D.G. Inxon. Dept. of Biology, University of Waterloo, 
Waterloo, Ontario N2L 3til t^current address; Dept. /.oology. University of 
r.uelph, Guelph, Ontario HIG 2lf 1 ) 

Obje ctives 

The project «as undertaken in 198!) as an integrated field-laboratory program 
designed to determine the impacts of copper and zmr. t-oiitamination on «hite 
sticker t Oatostoaua cowiersQni ] populations in several lakes in the Nanitouwadge 
"listrirt of "Jorthwm Ontario. The Manitouwadge lakes were selected because the 
levels of copper (13 to 15 ug/I J and zinc 1209 to 253 ug/lj wore slightly higher 
Ihrtii fanadian water quality guidel int-s . The water quality guideline approach is 
Iwsnd on the assunption that lakes of similar tharacleristics exposed to sinilar 
crancentrations of netals will exhibit siwiiar effects, k previous study 
conducted in the mid-1970s had docuBfnted changes in white sucker populat Ions 
exposed to levels of copper (12 ug/I) ami tuic (2^5 ug/1) tMcFarlaop and Franzin 
IuTm) close to those identified at the Manitouwadge si^e. To test the impar's of 
netals, .ind the water ijual ity guidel ine approach, »e sei out to examine the 
white suoker popiilatlonR in tli^ Hanitouwadge district, 

Our original objectives were to IJ examine the growth -ind reproduction of 
whit-e sucker populations in the Manitouwadge chain, 2; to examine the larvae 
Oi'tginnting frtim the contaminated sites for evidence iif df;i:! imal ory ch.inges and 
i) to develop a protocol fur identifying the impacts of chemical contamination 
on fish populations. S-imples were col lected from six lakes in the Manitouwadge 
chain over the period of I'iS^-l'iS?. White sucker were evaluated for evidence of 
impact on growth, reproductive perfonnance, larval survival tuui tolerance of 
l.irvae to copper and zinc exposure. 

■Sumwor y 

White sucker reached the age of maturity between ft and ft years of age at all 
sites, and until 6 yi>ars of age there were no differences in length or weigtil of 
fish from control and Manitouwadge (contaminated) sites [riiniki ttrick and Dixon 
i>>aOa). After maturit) , fish from contaminated sites were signif iitantly smaller 
and shorter than those from (ontro! sites. In addition, fish from contaminated 
lakes also exhibited decreases in fecundity ^md egg sir-', failed to show 
significant increases of fefundity with agp and exhibited an increased incidenne 
nf spawning failure (Hunkittrick and riix-jn 19B8a,h). 

Examination of the reproductive perfgrmance did not detect differences 
between white sucker collected at Conlaminat ed and uiintrul sites (Muiikii trick 
•inil Dixon wsehj. There were also no differences in the ferti 1 1 Jai ion rates of 
natural ly-ferti I i/ed eggs and in cross- fert i It lat ion trials the metaI-fiX[>o«ed 
males performed better than control mates in fertilization Trial'i with control 

Larvae hatched from eggs collecied at contammatetl sites were smiiller, 
developed a* a slightly increased rate, and exhibited poorer growth and survival 
than larvae from control sites (Munkiltrick and Dixon l*)8Sb). These changes 
were evident despite the fact Ihal the contaminated eggs were fertiliEeri and 
hatched in (lean water, and the differences are consistent with both the 


pheiiuniPTia of decr^^aseil female reprodat t ive commitnient and the vertical 
trntisuission (mother lo nffspritig) of coiit.dminont residues. 

Ttie trtiliire of fein-ile whitt sucker to grnw significantly after maturity, 
Btid the liec reused energpt ic coMmilBient t.D reprodijctiori suggested That t hf food 
l»asf in the contaminflted lakes was timitiiig thi' performance of the female white 
sucker. Female fish Bxhi luted dt'cri^'is.ed muse !e 1 ipid levels, decreased serum 
lipid levfls during tht' postspownirig period and on opjMrent decrease in Msterul 
lipidti 'Junng the autumn. There »erp on effects of collection site on body 
stnrHS of liver gl jrogen , I iisr lipids, serum triglycerides., total seruin 
cholesrerol (Munkittrick and Dixon I9l!fla) or brain amine levels (Hunkittrjck et 
al, 19fl8a). 

larvae showtd sigiii I icant chaiigei. in loltrarice and resistance to copper and 
zinc »ilh age and metal resistanije peaked al the tune of the ouse' of liver 
f unci ion jng. Larves frum contiuniuat ed eggs ahn«>;d increased resistance and 
tdleralite to uiiteiljonie coppei- during the ptfriodi of endogenous nutrition, 
dpspite II. u firi that the fggs were not pre-exposed to exogenous metals 
tHunkittrick and Diiton l^HHh) . Tiii: effect was no* seen in larvae at first 
fnedlng, at ages older thuii U d ofler 1 h-^ unsiit of fee'ling, sugg'isting that the 
cliange w«s n<it gen^tit. in origin. 

The effetl=i wos aIso absfftit in larvai- hatched from control eggs fertilized 
Kjth ipi-rm token frnm t»rtlcB at ront aminated sites. Fertilisation of eggs 
collrc.ted al the contaminalpd site with iiiilf rollected from control males 
yield ed larvae whose it'lerancH and resistance profiles could not be 
diftt iiigiiishtrd 1 rtini the Manit ouwidge larvae '.Munlil trick and Dixon I'VSfii:) . The 
far I thill t hr increase in loli*rante n^s as sot ia ted with eggs from t lie 
r.ontaiBiiialed '■\<f /md not thp milt suggests the presence of a maternal yolk 
factor associated »itb ini:r*fased resistance and tolerance of larvae to copper. 
Th« factor appear', t^ be mi'tal ri!.><idues transferred in the yolk, and no 
difffrences were detected m '■gg met al lolhionem residues het wefin control and 
i:oMaiiiiiiated sjt'.'s IMunki t tt ii k jiod Lixon 14Bfii J - 

Eggs from thu Man! touwadge *i te were significantly smal ler than control 
«;gg.s. 'ind naiuroUv-fert ihz^d r-ggs collfcted from the contaminated spawning 
sites exhibited a further d-'ci-ei^p tii egg si/p and an increase in deformity rat>: 
not evident in runt .iiiiinat ed eggs manuall\ fprtilized in control water 
[Muiikittrick and fUxon l')8fir). Iiicuhation trials involving the placement of eggs 
in hli^eams flowing oul "f 'h- failings area ret^olted in u decreased egg size and 
tolpranri' in i-oppi-r and .^n incr«aspd dt-formity ratp (flunkittnck and Uixon 
I'MBfj, Both 'Changes werp assfujiated with thi; ioflux of mi-tals during the 
Wat er-hardi-n J ng pi ori-SB . 

Tltf rii«;t rlhut ion of mel«ls m *hil(? sucker tissues wus monitored, and 
elevdtirnis m hot h i np]>er and iint residues were identified in liver, kidney, 
gill and grinadiil tissue iMilli^r et il . I'^ftS). Muscle levels of zinc were 

atftually -iigiuf h am ly lower at L-ontamj nai ed sites than at controls. There was 
■■A'iden't- uf iin?la! uptakt- liom thi- di<-t and the concentrations of metals in gut 
contt-nl-. eX'.eeded 'tfJ'D pjim In ^iid l^Ull ppMi Zo . Analysis of setllinent metal 
mnii .ui'r.jl ions slioneil eievai jun.s in lioth Co and 7.n ^t ri,ntamiriated sites iMillfr 
-\ <il. vmi; Munkltirii.-k -t .il. t9Bflb).. 

Addiii.irril fork showh that sfveral major food groups are missing from the 
■ipdimen! s ni font .^mitiated siti-s tHunkittrick et at , r!f8flhj , and prf^viou.5 work 


5ngg«>sts that sirdiments undpr water deeper than 5 m may be incapable of 
supporting siacroinvertehrnte fauna tG^nnan l"*?!: Pugh ^nd Molci W86J. Analysis 
iif bpfiihic samples colUctpd from near-shnrt- areas ( < ^ hi depth) at the 
rrontamiiiated sites indicdted n decreased abundance or absence of po! lution- 
sensitive groups such as ephemeroptera, plecoptera. odonata, hirudinea, unionid 
i.Ums, gastropods, aaiphipods and aquatic beetles (Mimkittrick et al . 1988) . 
Frtuiia at the contaminated sites was donina'ed by chi ronomids and other 

White sucker stomach contents showed marked differences between the sites 
iNimkittrick et al. ISfiSh; Munkittrick and Dixon waed). Fish collected at the 
control sites had an average of 7.8 organisms per stowach -ind were dominated by 
epb erne ropte ran larvae. Stomach samples collected at Manitoitvradgt' yielded «n 
ftveragi; of <i9.8 organisms per stomach and were domitiated by chironomid larvae 
t24 per stowach) (Munkittrick et al. I'JfiBb; Munkittrick and Dixon l^Sfld). The 
absunce of ephemeroptean larvae it contaminated sites and a decreased chironomid 
density in the sedimfnts would resytt m a psarked decrease in feedinu. efficiency 
■1' the rontaninated sites. 

Changes in bincliemical paraneters indicative of chronic stress were non- 
uxistant on inconsistent (Munkittrick and TJixon I'JftSi; Munkitttick et al. 
I'^flSaj. Effects on the growth, fecundity and lipid status of white sucker tould 
'le attributed to nutritional deficiencies related lo the decreased food 
ilnindaitcf and density at LnntaHiinated sites, which rould be related to the 
itirri'ased sediment metal levels. Sediment metal burdens have declined 
substantially since the late im60"s (Oermati W'TJ. Direct effects of the metals 
were .letectcd on the lat-vae hatching fn.m t-gys collected at contaminati-d sites. 
Pvidence for direct effects on egg sine and larval deformities were related to 
incrertsed metal burdens in the eggs. Tliis im:rease could be related tn both the 
'■iili-y of metals during fhe water-hardening process at corilauiinated sites and the 
vertical transmissiun of metal residues from the fenwie lbroi.igh 'he ynlk. 

Ill summary, white sucker cnlleiited from the Manitouwadge site showed a 
rtecreatied growth rate and fecundi ty, with nu apparent changes in niean age, 
cutirti t ion factor or egg f erti 1 izat ion abi 1 ity. Tins dties not compare we! I with 
McFarlane and Franzin's (1<I78J findings of increased growth rate and fecundity, 
decFeased m'-an age and di-creased repro'iufitive iierfonnance. This is surprising 
siiue twtli lakes were exposi^d to similar waterhorne metal concentrations, and 
w.-re of similar size and water h.irdness. The apparent inability to generalise 
ecosystem responses to seemingly identical cheinicijl stressors complicates, our 
abilities to jwedict ecosystem responses without details field sampling. 

There is a need to develop simple and inexpensive methodfi to follow fish 
population responses to environmental degradation or lake restoration. In 
addition to field and laboratory testing, a framework was devirluped -is a ample, 
cost-effective, rapid mechanism for assessment of toxicant impart on aquatic 
environments (Munkittrick and DivRti 19B8d,e,f). The framework. Population 
Indicators of Sublethal lontaminant Effects on .Surrkers (PISCES), separate.^ 
response patterns based on popiilat i un cliaract erist i cs . The framework is ati 
adaptation of Colly's (l^e^i) dfscnptions of fisheries exploitation impacts on 
fish populations. 

The status of fish populations is a reflection of the overall condition of 
the aquatic environment. The framework assunes that changes in th'' diiath or 
birth lates of fish populations, or alterations in the availability of food or 


habital are associated wil h charatiterist ic responses of sucker popiilatioiis 
(MUnkJttncl: and Dixon lySBe). The responses have been grouped into five main 
put terns basi!'] on population tharacterist ics sucli as mean ige. (ecundity and 
condition fnftor. The pattiitns correspond to direct effects on ddults 
(«)£plo!tationj , rpcruit.Beni failure, miiJtipli; stressnrs, food Ijnitation and 
niche shifts. Populations whirh are gnmum, [eproducmg oi surviving at rates 
which rtre indi st iiigtii sh«ble from a reference tcpntrolj population are considered 
frpp from adverse chemii;nl effects. 

The rtpplicution of PISCFS to this study, and Id several previously 
published dat.t sets, showed that white sucker popu Litions responded to 
envirnnmental stressors m predictahle piitterns (Munkittrick and Dixon I^88eJ. 
The system can also be applied to populations ist other species of fish, 
ini;1udlng salnonids, percitls and centrarchids tMunkittrick and Dixon I'SHSf J . 
However, there art- limitations associated with the selection of sentinel species 
and saiiTpling site^ which niujit lie taken into ronsideratton (Munkittrick and Dixon 

The use of fhe framewfirk is litniteij hy the selection and appropriate 
■sampling of a contpflralile control population (Hunk itt rick and Dixon l^aSe) . 
Additional 1 inn tat ions include the lack of dose-response sensitivity and 
predictive ability. In spite of the limitations, the f'ISCF.S dpproaiih does offer 
researchers looking ^t I Wild ^ites an early indication of the site of stressor 
impact on an ecosystem and provide tiselul inforniJit ion for the design of 
sdupliiig schedules and derivation uf useful, testable hypotheses, Wlien changes 
are not correctly predicted, the use of contrjist ing, generalized response 
pa'teriii can rict In direct and foi.'u<; research efforts on crucial areas impaci-ed 
by ch-inging rr,nditions. The .ivoi labi 1 ity of historical data sets and inf onnation 
from angler or fisheries harvests allows tlie PISCES system to be easily adapted 
for uonitoriiig purposes [Munkittrick and Dixon 19BBe). 

This study was funded bj nn Ontario Ministry of the Environment, 
EiiviroitMental Research i^ranl, We ary grateful io Dr. C. •ievitle (Rexdale) for 
her assistance rlnnng the slud\ . Me are rtlso giatefnl to Noranda for their 
cooperation, <-spei. lal ly M. <ipeyer (Hnranda We6««rt.h, Montreal! and L. Urhanoski 
tS'oranda OEiVO, lanit nuwadgej . We also wish to thank the Ontario Ministry of 
Vatnral Resourres for their cooperation, espe^ lal ly D. 1 ove tHanitouwadge J and 
I,. Melnyk-Ferguson ITerrai.e Bayj . 


tolby, P.J. lv8A. Appraising tiip status of fisheiies: rPhabi 1 it.ition techni(|ues. 
p. 2:J:)-257 fii V.H. Cairns, P.V. Hodson and J.O, Nnagu teds.). Contami nant 
Eff ects on Fishe ries. *dv. Environ, Sci. Technol. Vol. Ih., M.K 1^71, Tffeds of and mine wastes on the chemistry "tid ecology of 
lakes in the Manitoiiwadge i;hain district of Tliunder Bay. iint. Water Res. 
ritnmi. Spec. Repi . , Ontario Ministry of Environment, Thunder Bay. 

McParlane, O.A. and W.G. Franzin. 1^7H. Elevated heavy metals: a stress on a 
popul at ion of whi le surkTs Cat o si nm us coMMerson^, xn Hamel 1 Lake > 

Saskatchewan. J. Fish. Bes. Bd. Can. 35: 963-'>70. 

Miller, p. A., K.R. Hwnkittrick and D.G. Dixon. The liistribution of copper and 
zinc in tissues of white sucker [ Catoatowus coraaersoni ) froa control and 
metiil-contaniinated laXeii. Subnitted to Environ. Pollut. 

Mimkittrick. K.R. and D.G. Dixon. 19B8a. Growth, fecundity and energy stores of 
white sucker [ Catostomus commeraoni ) exposed to elevations of copper and 
■/.int. Can. J. Fish. Aquatic Sci.: ^5: 1355-1365. 

*(unkittrick. K.R. and D.G. Dixon. I'^aUb. Evidence for a waternal yolk factor 
associated with increased tolerance and resistance of feral white sucker 
(Catostonus cQniaers gni ) to w<iterhorne copper. Ecotox. Environ. Safety 15:7- 

Monkittrick, K.R. and D.G. Iiixon. l^uflc The effects of natural exposure to 
copper and zinc on egg size and larval copper tolerance in white suck«r. 
Submitted to Ecotox. Environ. Safety. 

.Mimkittrick, K.R. and D.G. Dixon. l-iSfld. In sttii assessment of copper and zinc 
impacts on white sucker populations of the Mini t ouwadge chain of lakes. 
Ontario Ministry of the Environment, Project 193 RR/.T3I PL, iO-i p. Liason: 
Dr. C. Neville. 

Mnnkittrick, K.R. and D.G. Dixon. t98ae. The use of white sucker populrstions to 
assess the health of aqua tic ecosystews exposed to lnw level chemical 
cnntaninatlon. Can. .J. Fish. Aquat . Sci. : IN PRESS 

Munkittrick, K.R. and D.G. Dixon. 14A8f. A holistic approach to ecosystem health 
assessvent using fish population characteristics. Submitted to Hydrobiologia 

Mimkittrick, K.R., F.J. Martin and D.G. Dixon. ISflfirt. The potential use of whole 
tiram amine and indoleamine concentrations as biochemic-il indicators ot 
i;orta»inant stress in fish. SubdiLtted to Co»p. Bujchpm. Physi'il. 

Miinkittrink, K.R., D, Barton, D.G. Dixon ,ind S. 8uni>. l'>S6h. Evidence for n 
niche shift by white sucker exposed to nixed iietal mining wastes. 15th 
Atjuatic Toxicity Workshop, Nov. 2a - Dec. 1, l^fifl, Montreal. Qufbec, 

Pugh, D.M. and L.W. Maki. 198t). Berithic corriBiuiiities in thf Manitouwadge Lake 
fhain, 1975 . Tech . Support Sect ion, Ontario Ministry of Environment, 
Northwest Region, Thunder Bay, Ontario. 


Effects i)f nixcd nt^tdl nining wustt- on white sucker popitlatioDs: deve I opsent of 
a fraaewurk to describe fish (xipuldtiun responses to cnvironaental change. 
K.ft. MunkJttrick und V.ii. Uivtm. Dn^-pi. ol" Biology, University of Waterloo, 
Waterloo, Onlirio N2L 3G1 ('currant address: Dept. Zoologyi University of 
tiuelph, Guelph, Ontario N'tG 2VU 

O bjectives 

The projRct was undertaken in Tiiflfi as an integrated field-laboratory program 
designed to d«t ermine tlie inpacts of copper and zinc tiontami nation on white 
jj.ucker [ Catosi hitids c uwimr-soiii ) pftpiilat ions in 5'-veral lakes in the ?1anitoitiradgK 
district '■if Niiitherii Ontnrio. Vhe 'l.iiii tfjTiwadge laVes wm sn'^ctod bttrause the 
iKVfi)^ nf rtippnr 1,11 to 1 S >ig/ 1 J and niiit UOi* to 2'i^ ug/l) *ere slightly higher 
ihiin I'rtnsdiaii water qiialitj giiidi"! i iins. Tli« water quality guidplllio rtppmiKh is 
lw-11-ii '10 th'- I'-sujnpt ion that Irtkes of similir characteristics exposed to similar 
concntrfltions "f inftals will exhibit limilar effei.ts- A previous study 
ifundiK.'tprl ill I lie (ttid-l')7iK hn.l docjuiieiileil changes in white sii'.ker pnpiilrtTiuns 
puifl)^^^ \n l*>v.?ls of t.opi'er il2 ng/lj intt i'lnc tJ'iS tig/l ) (MrFarlaiie /uiii Frotiitiii 
I'i7i!> rliisf 111 llio'ie ifl^i-iit 1 f "-d «t the Iflnitnimadg- sil". To test the impa':ts of 
mi' tils, '11 id llie Slit fM' qij'i I i ly giiidul nii- rtppl', Wf; s«t nut to PXamine »he 
t>hite Slicker pupijlatiuiis in the Innitoiiwadge distnci. 

i^iir original i>lijet'i ivea wiTP to 1 ) eX'iniine the growth and teprodiiction of 
tahi tf vnclier popvildl ions in 'he flnnit ouwiilge chain, 2) to fixamiiip the larvae 
origMiaMng from thi? 'inii ■imindte'l sjlr?s for evidi-nfe nf rtccl mattiiy changes and 
1) to devflnji a jiclocol fm itlnnti fying tlt^ of rhemicol contaminil ion 
nn fiKb poiJiilnt lOTis. SiiBtili"^ w;re Lollfjijted friwi six lat-es in the Manitoumadge 
ch.-iin iivi-r t lif pmod of l'*B')-l'<B7. White snrksr wei e evaluated for evidence of 
iiii|>.ii'i on growth. ri>pi-odiicl nw p*?rf oriMnoP, larval surx-iva! and tolerance of 
Uiv.i" ti, I itpjwc ^ii'J / f\|«.iure., 



White Slicker reached the age of maturity between « and 6 years of age at all 
sites, and until 6 years of age therp «ere no differen<~es in length or weight of 
fish from control and Hanitouwadge (rontaainated) sites iMnnkittrick and Dixon 
waSn). After maturity, fish from contftijiiiiated sites were significantly smaller 
and shnrter than those from control sites. In addition, fisli froB contaainated 
lakes also exhibited decreases in fecundity and egg size, failed to Bhow 
significant increases of fecundity with age and exhibited an increased incidence 
of Bpawning failure {Munkittrick and Dixon l98Ba,b). 

Exaaination of the reproductive perfirrmarice did not detect differences 
between white sucker collected <it contaminated and control sites l^fllnki ttrick 
a\,ri Dixon 19«8bJ. There were also no differances in the fertilizatioti rates of ly-ferti lizBd eggs and in cross-fertilization triaU the netal -exposed 
mains pertomed better than control pales in ferlil izition trials with control 

Larvae botched froie eggs collected at cuntaninated sites were smaller, 
(Inveloped at a slightly increased rate, and exhibited poorer growth and survival 
thfHi larvae from control sites (Munkittrick and Dixon l<)8&b) . These changes 
were evident despite the fact that the cuiitaMlnated eggs werft fertilized and 
hatched in clean water, and the differences ar<> consistent with both the 
phi.Tiowena nf rfecreased female reproductive comimtment and the vertictil 
transHissJon {mother to offspring) of contaminant residues. 

The fai lure of female white sucker t<i grow significantly after maturity, 
and the decreased energetic commitment to reproduction suggested that the food 
base in the contaminated lakes was lintting the performance of the female while 
sucker. Female fish exhibited decreased muscle lipid levels, decreased serum 
lipid levels during the postsiiawiiing period and an apparent dscrease in visceral 
lipids during the autumn. There were no effects of collection site on body 


stores of liver glyt:igen, liver lipids, serum t. rigtytendes, totnl serum 
cholesterol (Munkittrick and Dixon 19fl8a) or brain amine levels (Munkittrick et 
rtl. 198aa). 

Larvne showed significant changes in tolerorice and resistance to copper and 
zi»c wjlh age and metal resistance peaked at the time of the onset of liver 
functioning, l.rtrvap f rntn contaminatGd eggs sho'ted increased resistance and 
tolerance to watertiomp ropper during the periods of endogenous nutrition, 
despite the fact that the eggs were not pre -exposed to exogenous metal s 
(Munkittrick and Dixnri 1988b) , The effect was not seen vn larvae at first 
feeding, at ages older than M d after the onset of feeding, suggesting that the 
nhange was not genetiL- in origin. 

The effects was also absent iii larvae hatched from control eggs fertilized 
with spi'rm taken f roni males at contaminated sites. PfrtilLzatmn of eggs 
ml 1 ec ted at the containinat ed site with milt collected froD control nales 
yielded larvae whose tolerance and resistance profiles could not be 
distinguished from the Manitouwadge larvae (Munkittrick and Dison VJeSc). ThR 
f fli.l thai t he increase in tolerance was associated with eggs from the 
t ed ;^itp and riui t h*^ milt suggests the presence of a maternal yolk 
factiir dssutiated wi t li increased resistance and tolerance of laT-vae to copper. 
The factor appetrs tn be metal residues transferred in the yolk, and no 
differences were detected in egg metal lothioiiein residues between control and 
Rflnt'^tiinated sites <Hunkittrick and Dixon I-ieBcj. 

Eggii from the Mum louitadge site were significant ly smal ler than control 
pgg>>, and n'lfiiral ly-r'trr ' 1 1 ized eggs col Icted from the contaminated spawning 
sites exhihited a further decrease in egg >;iie and an inciease in deformity rate 
iioL evident in convaminated eggs manually fertilized in control water 
tMui.ikitt ri;':t< and Pix.on WOtt^}. Incuhot-U'ti trials involving the placement of eggs 


in streams flowing out of the tailings area resulted In a decreased egg size and 
tolerance to copper and an increased deformity rate (Miinfcittrick and Dixon 
I968cj. Both changes were associated with the influx of netals during the 
water-hardening process. 

The distribution of netals in white sucker tissues was aonitored, and 
elevations ill both copper and zinc residues were identified in ! iver, kidney, 
giil and gonadal tissue (Htller et al . 1486). Nuscle levels of zinc were 
ncturtlly significantly lower at contaminated sites than at controls. There was 
evidence of netal uptake from the diet and the concentrations of metals in gut 
contents exceeded WO ppm Cu and 1200 ppm Zti. Analysis n£ sediment, metal 
concentrations showed elevations in both Cu and Zn at contouinated sites (Miller 
et al. 1986; Hunkittrick et al. 198eb). 

Additional work shows that several major food groups are missing from the 
sediments of contaainated sites {Munkittrick et al. 1988bj , and previous work 
suggests that sediments und^r watnr deeper than 5 m may be incapable of 
supporting macroinvertebrate fauna (German 1S»71; Pugh and Maki 1986J. Analysis 
of benthic samples collected from near-shore areas (< 3 m depChJ at the 
contaminated sites indicated a decreased abundance or absence of jwl I ut ion- 
sensitive groups such as ephpmeroptera, plecoptera, odonata, hirudinea. unionid 
clams, gastropods, amphipods and aquatic beetles (Munkittrick et al. 1988), 
Fauna at. the contaminated sites was dominated by chironomids and other 

White sucker stoaach contents showed narked differences between the sitns 
tMunkittrick et al . 19aeb; Munkittrick and Dixon 19ead;. Fish collected at the 
control sites faod an average of 7.8 organisms per stomach and were dominated by 
rtptiemeropteran larvae, Stomarh samples col lee ted at Mani touwadge yielded an 
Average of 't9.8 organisms i>er stomach and wpre dominated by chironomid larvae 
(29 per stomach) (Munkittrick et al. 1988b; Munkittrick and Diiton I9e6d). The 


absence of i^pheBiprnptean larvae at contaBinoted sites and a decreased chironoieid 
density 50 the sediments would result in a markHd dccrensi? m feeding efficiency 
at the contfiminated sites. 

Cltange^ in biochemical paraiieters indicative of chronic stress wei^ non- 
en is ta.iit un inconsistent (Munkittrick and fJixcin l9B6al Munkittrick et at. 
l^SBaJ. Effecfs on the growth, fecundity and lipid status of vfhite sucker could 
tie attributed lo nutritional dsf i eienrics related to the decreased food 
abuiitlruicp rtiid liens ity nf cnni iimiriat*;d sites, «tn th rniild be related tv the 
increased sedij«ent metal levels. Sediment metal burdens have declined 
substantially since the late Ti60's (fierman T>71J. Direct effects of the iietols 
nerc detected on the larvae hatching from eggs collected at contaminated sites. 
Ev id^nce f nr direct ef f'-rt s dii t-gg size and larval ileformities weri; related to 
mr.tPfissd (m^ burdens in the t^ggs. This ini^rease could be related to both the 
entry if lopiaU during ihi> »ai er-hardeoing proiess at contaisinoted sites ind the 
v(*rtical t ransntssion of setnl residues from the female through the yolk. 

In summary, while sucker collected from the Manitouwadge site showed a 
decreased growth rate and fecundity, with no apparent changes in niRan age, 
condv tiort fact-ir or i"gg f ert i I i ?ot ion rtbi lily. Thi s dues not compare wel 1 with 
McFiir-lniiP ^iiiM Fran?, in' s t. !'"•*) f imHrigs of i iicrRfised growth rvi' h ;irid fecundity, 

dec^t^asM ntran age and decreased reproductive performance. This is surprising 

j.inf.e both lakes were expused to similar waterborne ineta! concentrations, and 
wer^ of siaii lor si?,p and water hnrrtness . Tlie apparent inabil ity to gen«ralize 
wfiosy.stpm rf-sjiotises in seemingly identical i-hemiral stressors coBpl icates our 
obiltties tn pri-rlift ff:o'-ystpni responses withou' detailed field sampling. 

There is .1 need I 'i develop simjile and iiiexpeiisi vp methotis to loll'iw fish 
)>o|Hilation responses tii environmental degradation or lake restoration. In 
addilinn fti I ield ami Iribofritorj testing, n frainework was developed as a simple, 


cost-effective, rttpid Bechnnism for assessment of toxicant inpact on aquatic 
environnents (Munkittrick and Dixon l<iQed,e,f ) . The t'raaework, Population 
Indicators of Sublethal Contaminant Effects on Suckers l PISCES J, separates 
rnsponsR patterns based on population cbaracterist ics. The franework is an 
adaptation of Colby's (.I'JB^) descriptions of fisheries exploitation inpacl s on 
fi5li populations. 

Tlie status of fish populations Is a reflection of the overall conditiin of 
fbe aquatic environment. The fraaework assumes that changes in the death or 
birth rates of fish pcipulations. or alterations in the availability of food or 
habitat are associated with charocterist ic responses of sucker poputiitions 
(MUnkittrick and Dixon IS^eSe). The responses have been grouped into five- main 
patterns based on population characteristics such as mean agei fecundity and 
condition factor. The patterns correspond to direct effects on adults 
tesploitation), recruitment failure, multiple stressors, food limitaticin add 
niche shifts. Populations wlii'.h are growing, reproducing or surviving at rates 
■ hich are indistinguishable from a reference (contrci) population ai e cons.idered 
free from adverse chemical effects. 

The application of PISCES to this study, and to several previously 
published data sets, showed that white sucker populations responded to 
environmental stressors in predictable patterns (Munkittnck and Dixon l^SSej. 
The system can also be appi led to popu I a' ions of other species nf fish, 
including salmonids, percids and centrarchids IMunkittrick and Dixon ivfiflf). 
However, there are limitations associated with the selection of sentinel species 
and sampling sites which must be taken into consideration iMunkittrick and Dixon 


The use of the framework is limited hy the selection and appropriate 
sampling of a comparable control population (Munkittrick ond Dixon l<)B8ej. 
Additional limitations include the lack of dose-responso sensitivity and 


prertirrtive ahility. In spite of the 1 inutrttions, the PISCES approach does offer 
resedrchprs looking at field sites an early indication of the site of stressor 
iopai:t on an etivsysteai and can provide useful a.nformat ion for tlm design of 
sampling schedules and derivation of useful, testable hypotheses. When changes 
are not correctly predicted, the use of Contrasting, generalized response 
patterns can act to diriict and focus research efforts on crucial areas impacted 
by (^hanging conditions. Tlie avai ifibi lity at historical data sets and information 
trom angler or fi&heries harvests allows the PISCES systetn to he easily adapted 
for rNDnitonrig purposes (Munkittnck and Liton I96ee). 


This study was funded by an Ontario Ministry of the Environment, 
Environmental Research Grant, We are grateful to Or. t'. Neville [Rexd^Ie) tor 
Imr flfisi^itanre di-iring the study. We are also grateful to Noranda for their 
cooperation, especially t1. Speyer [Noranda Aesearch. Montreal) and L. Urbanoski 
(Noranda GKCO. ManitouwadgeJ. We also wish lo thank the Ontario Hinistry of 
Vflliiral Resources for their cooperation, especially D. Love (Msnitouwadge) and 
I. . Me I iiyk-Fergusoi] (T'^rr.jc*: BayJ . 

R eferenceH 

Colby, P.J. \^Sit. Appraising the status of fisheries: rehabilitation techniques. 

p. aas-i.'}? In V.W. Cnims, P.V. Hodson and J.O. Nriagu (eds.j, Contaainant 
Effects on FiBherii-fi , Adv. Environ. ,Sci . Technol. Vol. Ih. 

German, M.I. W?!. Effects of acid mine waFites on the chemistry and ecology of 
lalies in thp Mail) touwadge rhajn district of Thunder Bay. Ont . Water Res. 
Conan. Spec. Bept . , Ontario Ministry of Environment, Thunder Bay. 

NcFarlane. G.A. and W.G. Franzin. 1976. Elevated heavy metals: a stress on a of white suckers Ca toat pmiis comjersoni, in Hamell lake. 
Saskatchewan. .1. Fish. Res. Bd. Can. 15: 963-970." 

Milter, P. A., Munkittnck and fJixon. The distribut laii of copper and 


zinc in tissues of white sucker t Catostowius cownerson i] from control and 
metal-contaninated lakes. Suhoitted to Environ. Pollut, 

Hunkittrick, K.R. and D.ii. Dixon. IWda. Cirowth, fecundity and energy stores of 
white sucker ( (.'atostomus cowMersotii ) exposed to elevations of copper and 
/.inc. Can. J. Fish. Aquatic Sci.: «: 1355-1365. 

Munkittnck, and D.lJ. Dixon, isaab. Evidence for a natemAl yolk factor 
associated with increased tolerance and resistance of teral white sucker 
( Catostoaus coanersoni ) to waterborne copper. Ecotox. Environ. Safety 15:7- 

Ilimkittrick, K.R. and D.G. Dixon. t4a6c. The effects of natural exposure to 
copper and zinc on egg size and larval copper tolerance in white sucker. 
Submitted to Ecotox. Environ. Safety. 

Kunkittrick, K.&. and D.G. Dixon. 1486d. In situ assessaent of copper and ;tinc 
impacts on white sucker populations of thn Nanitouwadge chain of lakes. 
Ontario Ministry of the Environaent, Project 193 RR/331 fL, 209 p. Liason: 
Dr. C. Neville. 

Hunkittricki K.K. and D.G. Dixon. 1406e. The use of white sucker populations to 
assess the health of aquat II ecosystems exposed to low level chemical 
contaninotion. Can. J. Fish. Aquat, Sci. : IN PRESS 

Kunkittrick, K.R. and D.G. Dixon. IvSHf. A holistic approach tu ecosyaten health 
■issesment using fish population characteristics. Submitted to Hydrobiologia 

Kunkittrick, K.R., Fl.J. Martin and D.G. Dixon. I'vUNa. Xhp potential use of whole 
brain anine and indoleamme conceittrat ions as biochemical indicators of 
contaninant stress in fish. Submitti-d to Conp. Biochen. Pbvsio!. 

Muiikittrick, K-B., D. Barton, D.t5, Dixon and -S. Bunn. l^SBb. Evidence for a 
niche shift by white sucker exposed to mixed atetal mining wastes. 15th 
Aquatic Toxicity Wnrkshop, Nov. 28 - Dec. 1, 1986. Montreal, Quebec. 

Pugh, D.M. and L.W. ffaki. 1466. Benthic comnunities in the Hanitouwadge Lake 
chaiu, l'i7fi. Tech. -■jupport .Ser.tion, Ontario Hlnistry of Environment, Region, Thunder Bay, Ontario. 



R.P. I^nito and D.G. Dixon, Departaent of Biology, Universitv of 
Waterloo, Waterloo, Ontario, K2L 3G1, 


Cyanide (CN") is used by the mining industry in the 
extraction and concentration oE gold and silver from their 
respective ores. Both froth flotation and leaching utilize CN~ 
for solubilization and complexation. As a result cyanides are 
routinely present in nine effluents in considerable quantities. 
This situation has long been recognized as an environmental 
problem and has resulted in the establishment of an Ontario water 
quality objective for CN" (0,005 mg/L, as HCN) (Ontario Ministry 
of the Environment, 1984). 

A number of processes have been developed for the 
elimination of CN" from mine effluents. Cyanide is often 
conplexed with sulphur, either from sulphur dioxide or an 
inorganic poiysulphide, to form thiocyanate (SCN") . Although 
set!" appears to be much less toxic than CN~, there is relatively 
1 ittle scienti fie evidence to fully substantiate this 
observation. As a result, there is currently no water quality 
objective for SCN~ in Ontario and no sound data base to establish 

The 96-h LC50 values for SON" for freshwater fish range from 
50 to 230 mg/L (Speyer and Raymond, 1985; Doudoroff, 1976), 
suggesting that SCN" is substantially less toxic than HCN. Acute 
toxicity data for SCN" provides no information on the long-term 
effects of SCN" on the growth and reproduction of freshwater 


fish. Also, the toxic mode of action of SCN has not been 
identif iecl, but often leads to a sudden, violent death termed 
Sudden Death Syndrome {SDS) by Heming et al, (1985). 

The objectives of our research are to obtain sufficient data 
on the long-term sublethal toxicity of SCN" on the growth, 
metabolism and reproduction of freshwater fish to permit the 
establishment of a water quality criterion. An attempt will also 
be made to apply laboratory results to a field situation in the 
gold mining region of northern Ontario. 

Materials and methods 
Laboratory studies 

The laboratory portion of the study has been divided into 
two phases: 1) The lonq-tem sublethal exposure of rainbow trout 
fry to SCN~ to determine effects on growth and metabolism, and to 
characterize a syndrome of sublethal SCN" toxicity. 2) A life- 
cycle study on the effects of sublethal exposure to SCN" on the 
reproductive capacity of fathead minnows. 

The growth trial portion of the rainbow trout phase of the 
study has been completed and analysis of data is currently 
underway. The fathead minnow portion of the study has just 
recently been started. 

Acute baseline bioassays 

Sainbow trout (2 g) were exposed to various concentrations 
of SCN" for a 96 h period. Mortalities were recorded at various 
time intervals up to 96 h. At 96 h, fish were stressed by a 15 s 


pursuit with a hand held dip net, and subsequent mortalities were 
recorded 30 minutes after the application of the stressor. 

Chronic exposure studies 

Triplicate groups of rainbow trout (2 g) were continuously 
exposed to nominal SCN~ concentrations of 40, 80, 120 and 160 
mg/L for 16 weeks. Fish were randomly allotted to 60 L, white 
fibreglass tanks and fish weights were standardized to a 
coefficient of variation of <;3% the day before the commencement 
of SCN~ administration. Trout were pair-fed a practical trout 
diet (GRT-70) (Cho et al, , 1974) . Each treatment tank was 
randomly matched with a control tank within its block. Treatment 
tanks were fed ad libitum and the ration weighed, four times per 
day. An equal weight of food was then fed to the matched control 
tank for each treatment replicate. Trout were weighed every two 
weeks and the weighing procedure was also used as a routine 
stressor to measure the expression of SDS within each treatment 
population. Feed intake, mortalities and feeding behaviour were 
monitored daily. 

At the termination of the 16 week growth study, fish were 
anaesthetized in MS222 and killed by cervical dislocation. 
Length, weight, and splenosomatic and hepatosomatic indices were 
measured. Hematocrit and hemoglobin determinations were 
performed. Plasma samples for the colourimetric determination of 
total plasma cyanide-reactive substances (Lambert et al., 1975) 
and plasma thyroxine (T^) and trl-iodothyronine (T3) were 


obtained by the severance of the caudal peduncle. Samples were 
frozen and stored at -20°C until analysis. Liver samples were 
taken for the determination of liver glycogen and protein levels. 
Thyroid, liver, kidney, head kidney, gill, spleen, cartilage 
and blood smears were subjected to routine histological analysis. 

Field studies 
The Hemlo gold mining region of northern Ontario was 
selected as a site for the environmental health assessment study 
of the impacts of SCN~-bearing effluents on aquatic systems. 
Effluent effects on fish populations will be assessed by 
Donitoring growth (age versus size relationships) , reproductive 
capacity (size at sexual maturity, fecundity, egg size, 
hatchability) , histopathology of major tissues and biochemical 
parameters shown in the laboratory to be affected by SCN" 

Acute baseline bioassay 

The 96-h LC50 for unstressed rainbow trout exposed to KSCN 
was 250 mg/L. When mortalities due to the stress of a 15 a 
pursuit with a dip net after the 96-h exposure were included in 
the calculation, the 96-h LC50 was 180 mg/L. All fish that died 
after application of the stressor exhibited flaring of operculae, 
extreme muscle contraction resulting in a dramatic curvature and 
arching of the body, spasms, loss of equilibrium, loss of 
buoyancy control and changes in pigmentation, all signs 
characteristic of Sudden Death Syndrome (SDS) . 


Chronic exposure studies 

Mortalities and behavioural anomalies 

All fish exposed to 160 mg SCN~/L died by the end of the 12 
weeks. Many of the deaths coincided with the expression of SDS 
after bimonthly weighing procedures. Approximately 40% of the 
fish exposed to 120 mg SCN~/L had died by the end of the 16 week 
growth trial, with varying proportions of the fish expressing 
SDS after weighing. Mortalities at the two lower concentrations 
(40 and 80 mg/L) were minimal, except for one replicate (80 mg/L) 
which was situated near a corner of the experimental system, and 
as such, was often disturbed by laboratory traffic. Increased 
mortalities in this replicate may suggest an effect of an outside 
stressor on the expression of SCN" toxicity. Fish exposed to 
lower concentrations of SCN~ did not express SDS at any time. 

The major behavioural anomalies observed were irritability 
and skittishness. Fish exposed to 120 mg SCN~/L exhibited a 
behaviour in which fish swam rapidly in small circles for 10-20 
s. This behaviour ususally coincided with the fish being offered 
pelleted food and attempting to feed. 
Gross physical observations 

All exposure levels of SCU~ resulted in varying degrees of 
deformities in the cranial region of the trout. This condition 
was characterized by the small size of the head of the trout in 
relation to the body. The head was laterally and dorso-ventrally 
compressed. Operculae were often shortened or crumpled, exposing 
gill filaments. The severity of these signs appeared to increase 


with SCN~ concentrations. 

Pigmentation changes were also evident as the darkening of 
individual fish within tanks receiving thiocyanate. This 
darkening appeared to be transitory, as all fish in tanks where 
dark fish were observed would be lighter in colour upon 
subsequent observation. Liver soinatic and splenosomatic indices 
decreased in exposed fish and the livers of fish exposed to 120 
Big SCN~/L appeared pale and were friable. 

Blood parameters 
Fish exposed to SCN~ developed an ansniia characterized by 
decreases in hemoglobin levels and hematocrit. Further 
histological characterization of cell types in blood smears is 
currently underway. The decreased hemoglobin and hematocrit in 
conjunction with the decreased splenosomatic index are suggestive 
of a hypoplastic anemia. The magnitude of the decreases in 
hemoglobin and hematocrit increased at higher SCH" 
concentrations. Plasma total cyanide- reactive substances 
increased with waterborne SCN~ concentrations. Plasma samples 
are currently being evaluated Cor tri-iodothyronine (T3) and 
thyroxine (T4} levels. 

Tissue parameters 

The evaluation of tissues for various parameters is 

currently underway. Liver will be analyzed for glycogen and 

protein. Foutine observations during sampling suggest that there 

may be more visceral fat present in fish exposed to thiocyanate. 


Carcass samples have been frozen for routine proximate analysis 
of fat, protein, ash and moisture. 
Liver, head kidney, kidney, spleen, thyroid and cartilage 
have been sampled and fixed for histological analysis. 


Based upon preliminary analysis of data, long-term, 
sublethal exposure of rainbow trout to SCN" results in a syndrome 
characterized by decreased growth and feed intake, cranial 
deformities and a hypoplastic anemia. Plasma total cyanide- 
reactive substances are also elevated with increasing waterborne 
SCN~ concentrations. Sublethal physiological and morphological 
effects were noted at all concentrations of SCN~ tested, although 
there appeared to an increase in the incidence and magnitude of 
responses with SCN~ concentration. 

The sublethal mode of action of SCN~ is not well understood 
in fish. The sublethal effects of SCN~ in mammals are exerted by 
its antithyroids 1 activity, inhibiting the active uptake of 
iodine from the blood by the thyroid and acting to uncouple T3 
and T4 from their carrier proteins in the blood (Green, 1971) . 
Thiocyanate also actively competes for membrane transport with 
other halides such as chloride (Epstein et al. , 1975; Katz et 
al. , 1982) , and hence could be involved in ionoregulatory 
disruptions. How these potential modes of action fit into the 
sublethal toxicity syndrome seen in this study, remains to be 
determined, pending final analysis of data. 



Cho, C.Y., H.S. Bayley and S.J. Slinger. 1974. Partial 
replacement of herring meal with soybean meal and other 
changes in a diet for rainbow trout ( Salmo gairdneri l . J, 
Fish. Res. Board Can. 31:1523-1528. 

Doudoroff, p. 1976. Toxicity to fish of cyanides and related 

compounds; a review. Ecol. Res. Ser. EPA 600/3-76-038. 

Office of Res. and Devel . , Environ. Res. Lab., US EPA, 
Duluth, MN., vi;54 pp. 

Epstein, F.H., P. Silva, J.N. Forrest and R.J. Solomon. 1975. 
Chloride transport and its inhibition by thiocyanate in 
gills of seawater teleosts. Coinp. Biocheui. Physiol . 

Green, W.L. 1971. Mechanisms of action of antithyroid compounds. 
IE The thyroid: A fundamental and clinical text. Edited by 
Werner, S.C. and S.H. Ingbar. Harper & Row, New York. pp. 


Heming, T.A. , R.V. Thurston, E.L. Meyn and R.K. Zajdel. 1985. 
Acute toxicity of thiocyanate to trout . Trans . Am . Fish . 
SOC. 114:895-905. 

Katz, U., K.R. Lau, H.H.P. Ramos and J.C. Ellory. 1982. 
Thiocyanate transport across fish intestine ( Pleuronectes 
Platessa l ■ J. Membrane Biol. 66:9-14. 

Lambert , J . L. , J . Ramasamay and J . V . Paukstel is . 1975 . Stable 
reagents for the color imetrlc determination of cyanide by 
modified Konig reactions. Anal. Chem. 47:916-918. 

Ontario Ministry of the Environment . 1984 ■ water management 
goals, policies, objectives and implementation procedures of 
the Ministry of the Environment, Revised. Toronto, Ontario. 
70 pp. 

Speyer, H.R. and p. Raymond. 1985. The acute toxicity of 
thiocyanate and cyanate to rainbow trout as modified by 
water temperature and pH. Environ. Toxicol. Chem. 7:565-571. 






Q.M.Kirby.l.R. Smith", C. Thorn. H w. Ferguson and MA. Mayes 

Fish Pathotogy Laboratory. UrUverslty o( Guelph. Guelph, omario NIG 2Wi 

•Omario Mlnislry cpI Ertvtronment, Rexaale, Ontario, Canada, M9W5L1. 


Various liver lumors occur with increased frequency in several species of bollom-tJwellIng tish inhabillng 
ICKalions with indusirialty polluied sediments. In white suckers (Caiostomus cammetsoni) from the 
HamiRon Hartour region of Lake Ontario, preneoF^astc and neopiastic liver and bile duct neoplasms are 
well recognized but are much less prevalent in whits suckers from less-polluted control sites in Lakes 
Huron and Sinxxie. Potyctonai aniisenjm prepared againsl purttied hepaiic glutathwne S-lransterases 
IGSTs) trom wfiite suckers was used lor immunocytochemical demonstration ol GST expression in the 
various liver lesions in white suckers. GSTs are considered lo be Irrportant In detoxification ot polycyclic 
hydrocarbons (PAHs) and diminisfi Ihe mutagenicity ol t)enzo(a)pyrene (BaP). Ail liver lumors were GST- 
delicient in comparison vnth surrounding r>on- neoplastic liver. Also, these llsh had lew early preneoplastic 
loci compared with advanced liver tumors. White Suckers from Ihe western Lake Ontario region ^ad 
taster rates ol biliary excretion of BaP Ihan lish trom clean siies in Lake Huron. These findings suggest that 
environmental mutagenic chemicals such as PAHs. which can be normally detoxified by GSTs in these 
nsh, may play a role in the laier stages (mailgnarfl progression) of cancer devetopment in cells that have 
losi GST-dependsm resisiance mechanisms. This hypothesis Implies that tong-term exposure to PWHs 
may necessary to cause neoplasms In tissues that are tnitiaily resistant to them. 


The Hamilton Harbour region of Lake Ontario, like many other industrial sites in the Great Lakes, 
has a wide range o) organic chemical pollutants accumulated in the sediments (Harkiw and Hodson. 1986), 
In recent years, increased prevalences ot various skin and liver neoplasms have tieen demonstrated in 
bottom^Jwelling ttsh Irom ihe Hamilton Hartour region, especially in White Suckers (Cafosiorrws 
commersoni) (Sonslergard and Lealhertand, 1964; Smith and Ferguson. (966; Melcalfe ef al. 1987; 
Hayes «( a'-. B87; V. Caims. personal communication}. The geographic associalion of these tumours wHh 
increased industrial pollution suggests that Ihe affected tish may be exposed to carcinogentc pollutants, 
but the Implied cause-elfect relationship has not been clearly demonstrated. Because there are 
numerous posstoly carcinogenic agents m the sediments to which the affected fish are exposed (Hartow 
and Hodson, I9B8}. it is unl9(ely that any one dass of chemicals can be delinitively and exclusiveiy 


impricated by epidemiological studies. However, the abundance of various polycycltc arematic 
hydrocarbons (PAH) in the polluted environment (Hariow and Hodson. 1988). many ol which are known 
skin and hepalc carcirwgens for mammals and lish (Siaga eial. 1980, Hendncks e/ a/, 19B5I supports a 
reasonable suspicion ihai PAHs may play a role in ihe development ol ihe neoplasms observed in these 
fish. This suspicion is reinforced by eviderKe that similar neoplasms in other benihic lish m Puget Sound 
(Meyers el al.. IM7; Varanasi ei al. 1987) Lake Erie {Baumann and Harshbarger, (985) and Lake Omafio 
(MetcallB »f al. 1987: Dunn el al . 1988) are associated with increased exposure to PAHs. 

Our recent studies have addressed Ihs question thai PAHs might tje responsible (or Ihe liver 
tumours we have observed in While Suckers Irom Ihe Hamilton region We have been conparing ibe 
kinetics and metabolism o( benzo(alpyrene (BaP) as a typical carcinogenk; PAH in tish (rom the Hamilton 
region and from less polluted reference siles in Lake Simcoe and Lake Huron. These studies suggest 
thai lish Ifom the polluted sile have an induced ahdity to activate BaP in the Wer and to excrete H in Ihe bUe 
as glutalhkjne (GSH) conjugated metabolites. Furthemwre, preliminary evklence indicates that the liver in 
in White Suckers may be naturally resistant lo PAHs because of the normal hepalic glutathione S- 
If anslerases (GSTs) which are imporlani detoxification enzymes for PAHs in these lish. Developing liver 
neoplasms lose their normal GST enzymes and Ihereby likely become rnore sensitive to Ihe PAHs Ihat 
would tie adequately deloified and excreled by GSTs in rrormal liver cells Colleciively, our obsarvatrans 
support a hypothetical explanation for carcinogen-induced genetic alterations responsible lor maliflnanl 
Iranslormalion and cancer progression in cells that may have been originally quite resistant to PAHs and 
other carcinogens An imporlant aspect ol this emerging corx:ept is Ihat repealed exposures to high 
Closes of gerwtoxic caicinogeru may cause malignancies in restslani tissues, irxlivkluals or species, 

1 IDiSlribution and Excretion of Benzofajpyrene [BaP] 

Male lAffiite Suckers were captured from Sixteen Mile Creek near Oakville, Lake Ontario (polluted 
srte) and from Keefer's Creek. Lake Huron (reference slie) during their spring spawning migrations and 
were maintainod in laboratory holding lanks in clean well water. Fish were selected and given BaP (2 
mg/Vg] by gavage in a vehicle composed of distilled detonized water, DMSO ( 1 0%) and sodium 
deoxycholaie [1%). The BaP preparalion contained 50 tiCi/mg of ^H-BaP (New England Nuciear, Boston 
MA) as a radioactive tracer Treated lish were killed after 3, 6, 12 and 24 hours and subjeaed lo 
poslmonem examination, dunng which samples of bile (gall Dl^Jder), liver, muscle, blood, kidney, 
intestine, intestinal contents and gilt were cotlacled lor liquid scintillation counting (LSC) ol the ^H-tracer. 

In a second experiment. While Suckers Irom Oakville were heW lor 6 weeks under clean 
laboratory conditions belore BaP adminisiralton to determine If Ihe rales of BaP metabolism were altered 
when Ihe tish were no longer exposed to their polluted natural environment. 


Samples o( Wood and bile (40 nl) were sokjbiJized in i ml of Proiosol{NEN| and counted in 10 ml 
ol Aquasol (NEN). Samples ol tissue (100 mg) were digested and extracted ovemlghl in 2 ml 0.5N NaOM 
and 3 ml n-hexane on a motorized roiater (Varanasi efaf. 1978). An aliquot (1 5 ml) ol itie n-henane 
fraction containing non polar BaP metatxililies and the NaOH fraction (1 ml] containing polar metat»)iles 
were then counted In 10 ml of Aquasol. Concentrations of BaP and denvaliues were calculated as nmol/g 
tissue Irom liquid samillation counts ol viais after ovemigfit dark -acclimation and correction for counting 

2. Metabolism of SaP 

Samples ol bile obtained from fish given ^H-BaP were subjected to enzyme tiydrolysis and HPLC 
analysis to determine the proportbns ol B[a)P excreted as polar metaboilteE and conjugates. Bile In 1 ml 
of distilled water was extracted initially with ethyl acetate to remove non polar BaP (parent compound) 
The residual aqueous phase was divided into 3 equal pans brought to 9 mts using sodium acetate 
buffer, incubated with arylsulfaiase (Sigma Chemical Col.. Si. Louis. MO. 35 units'ml Incubate containing 
SO mM D-saccharIc acid 1 -4 lactone to inhtoit glucuronidasa activity) or ^-glucuronidase (Glucu rase. Sigma) 
or sodium acetate bolter (0.2 mM, pH 5,0, 37X. lor 24 hrs). Each sample was furitier extracied with ethyl 
acetate (2 x 2 nH) to remove less polar hydrolyzed BaP intermediates. Ail emracts and fractions were 
subjected to LSC to determine percentages of hydrotysible and nonhydrolysaDle polar metabolites. 
Samples ol bile (50 (ii) from fish given ^H-BaP 24 hours previously were anatyzed by reverse phase HPLC 
using a stepwise water/melhanol gradient on a C18 column (Biorad Hipore F1P-31Q: 250 x 4.6 mm) on a 
Blorad 402 HPLC system The rrxjb'le phase conditions were 0-30% methanol in 2 minutes. 30% 
methanol lor 1 5 minutes .30 -70% in 3 min and 70% methanol lor 1 minutes at a llow rate of i mtminute. 
The eluant was nwnrtored by absoibance (430 nm) and also by fluorescence (excitation 380 nm; 
emission 430 nm) by a Shimadzu RF 5000 spedroftuoromeler and collected as 25 ml Iraaions in a 
GHson Model 203 microiraciion collector. The distribution of ^h radioadivily tracer m Iraaions was 
determined by LSC 

3 Determinalton ol GST Activity and Expression in Rsh Tissues 

Samples ol liver, kidney, muscle, intestine and gill were also collected lor histopathologic 
examination (lixed in 10% formalin). Samples ol liver were also coHecied on ce, and homogenized in 3 
volumes ol 0,25 M sucrose buffer (coniaining 50 mMHEPESpH 7 5) lor delerminaliofi of hepatic 
glutathione S-translerase (GST) activity. Liver homogenales were centrituged at 100,000 g lor 1 hour to 
obtain cytosol (supernatant) Irom which GST activity was determined by CDNB conjugation rates (pH 7.0, 
aO'C) by the method ol Habig el al (1974), Pnjiein concentrations in cytosol were deiermtned by the 
Lowry method. 


GST isoenzymes were poriljed from liver cytosor tram normal While Suckers by aflinlty bindirtg to 
S-henytglulalhione-agarose (Sigma) and stepwise elulion wilt> 50 mM and 200 mM NaCI (to remoua non 
specflically bound proteins) and Ition 5 mM S-tiexylgluiatione (to elute glutathione -binding cytoslic 
proteins). The latter traction, containing approximately 60% ot hepatic GST, was analyzed by SOS PAGE 
urxJer reducing conditions and was found to consist o( 4 major protein subunits in ihe 26 kD molecular 
weigh! range corresponding lo reference samples ot pure GSTs from ral liver. The purilied GSTs were 
used to imnxjnizB rabbns to produce polyclonal amisenim specific lor all A GST subunrts in hepatic cylOSOl 
(bywesiem blot analysis). Tbis specific antiserum was used in routine peroxidase-antiperoKidase iPAP) 
immunocytochemical slainmg lor GST protein expression in (ormalm-fixod sections o( hvers from various 
Oakville and Hamilton Harbour lish with previously diagnosed nepatoceliular or bile duct neoplasms 


1 Distfibulton and Excretion ol BaP 

Fish from Oakville excreted BaP twice as fast as reference fish whan they were exposed to ^H- 
BaP (2mg/kg) within 7 days ol capture Irom ttie witd (Figure 1 ) . The vast majorrly of polar BaP meiatoliles 
were lound in the bile BaP-derlved radioactivity in muscle, bk30d,9tll. and kidney were negligitjie in tjoth 
populations (less than i % ot total) (Figures 2 S 3). large amounts of ^H activity were preseni in liver 
and miesline. Howevar. luilher analysis ol these latter samples by NaOH/n-fiexane exlracllon (see betow) 
revealed thai the intesline contained mainly unatjsorbed parent compound wtiereas the liver contained 
mainly polar metabolites ol similar types to those found in the bile 

The lish Irom Oakville exposed to BaP after 6 weeks in captivity excreted BaP at a lower rate similar 
lo mat observed in fish from the reference Site ai the lime of capture (Table 2), 

2. Metabolism of B(a)P 

The matonty of radioactivity in bile occurred as mors polar metabolites that coukJ not be extracted 

(rem the aqueous phase by ethyl acetate. The aqueous phase ol bile from Oakville and relerence flah 
contained some metabolites that were hydrolysed by aryi sullalase or 0-g)uCuronidase (Table 3). These 
experiments Indicated that Oakville Ush bad an increased proportion of sulfated conjugates 116.2 fotd ), 
but a similar proportion ol glucuromdes as reference fish (Table 3). Non hydrolysible polar melabolites, 
presumably containing gluialhione and olfter conjugates, comprised a substantial proportion ol the -H- 
BaP in bile trom each group. 

The disifibulion of fluorescent and ^H-labelled metabolites of BaP in hPLC analyses of whole 
unexiracied biie indicated thai fish from polluted and relerence sites had a similar complex prcifiie ol 
moderately polar and highly polar BaP melaboliles [Figures 2,3) No parenl (non polar! BaP was 
detectable in these bile samples. While there were minor, as yet uncharaclenzed, differences in some of 


the meiabolrtes. the observaiions were consistent wHh excretion studies (Table t) whk^ indicated an 
oweral increased rate oi BaP metabolism and eliminalion ol BaP in bile. 

3- Ofilermination of GST Aaivity and Enpression in Frsh Tissue 

Activities ol GST in hepatic cytosoi inDm While Suckers exposed to BaP In these expenmertts 
were subsiantially lower Ihan GST activities In While Suckers that had not been given BaP (Table 4), 
These observations are consistent wUh an interpretation ihat BaP metabolites compete with or inactivale 
GST in liver Fish trom OakvJBe had approximatetv one half the GST activKy wlien conpared with fish from 
Ihe reference site. 

immunocytochemical stains o( tissues from White Suckers revealed that GST proteins are present 
in substantial amounts in hepalocytes, bile duct epithelium. giK epithelium, intestinal epithelium, renal 
tubules and erythrocytes AJI hepatocellular and bile duct adenomas or carcinomas examined in li^ from 
Hamilton Hartxur or O^cvHle were matttedty deficient In GST proteins in comparison with surrounding non- 
neoplastic hepatocytes or major bile duels. Preneoplastic hepatocellular tesk^ns (foci of altered 
hepalocytes) which were rarely observed m livers ol fish with neoplasms, on the whole had similar 
amounts Of GST as dkl normal hepatocytes, but occasionally, GST defiaeni loci ol hepatocytes were 
observed. No preneoplastic or neoplastic lesions with induced GST were observed. 


These studies indicale that White Suckers from the industrially polluted western region ol Lake 
Ontario have evidence of an induced ability to metabolize and rapidly eliminate BaP in the bile This 
induction is transient and subsides when lish are held lor some lime inanunconiaminatedenviTOnmeni 
This evidence supports Ihe view Ihal these fish may be exposed to xenobiolics that induce various 
hepalic cytochromes P-450 artd deioxrflcatnn enzymes involved in elimination ol xenobliolics. A 
reasonable inierpreiatk>n ol these tindings Is that fish are Inlluertced by the polluted environment in 
western Lake Ontano, and that Ihis kifiuerKe helps them to excrete greater amounts of PAHs than can 
(ish Irom less polluted regions. 

The observations that White Suckers have numerous biliaiy rrjelaboliles of BaP are in accofdarwe 
wilh observations Ihat English sole from Puget Sound also excrete SaP by multiple pathways (VaranasI el 
al.. 1987). Our evidence suggests that corrugation with GSH by hepatic GST activity is a major 
delOJ(ification mechanism for BaP in While Suckers. Because these fish are capable ol rapidly excreting 
relatively large experimental dosages ol BaP. especially when they have lived near or in the Hamilton 
Harbourthat Is conlamlnaled whh many PAHs (Harlow and Hodson. I98B). it is reasonable lo consider that 
While Suckers are relatwety resistant lo BaP by virtue of their natural GST and other deloxHicalwn 
pathways. Such an interpretation is consistent with our ot>servations (Hayes el al. 1967) and those ol 


othOfs (V.Calms. personal comniunicatlon))(hat hepatic neoplasms are observed in fewer than 1 0% ot 
While Suckers tha\ have likely been exposed to the Hamilton environment lor many years. Moreover, 
preneoplastic loci ol allered hepaiocytes, of tt»e type considered to be initiated by PAHs and other 
genotoxic carcinogens in rodenis (Farber arxJ Sarma, 19B6) and fish (Hendricks ei al 1985; Meyers el ai 
1987), were rarely observed in these lish inspile Dl their presumably chronic exposure lo PAHs and other 
potentiail)^ genoioxtc carcinogens Accordingly, n is reasonable to conclude thai Whrte Suckers exposed 
to the HamiHon environmerrt are rather resistant lo any initiating etlecls ot Ihe PAHs to whtch they are 

Because there is an apparenily high rate of malignant progression (conversion) ol preneoplastic 
lesions to hepatocellular adenomas and carcirwrnas, these lish must develop an increased susceptibility 
10 tumorigenesis at some stage after the inttiaiion slep, lf> laboratory rodents, few ol the numerous loci of 
altered hepaiocytes initiated by brief exposures to genoioxic carcinogens actually progress lo malignancy 
(Farber and Sarma, 1986) By comparison, preneoplashc epidermal papiltomas initiated by PAHs m mice 
exhibit a high rale of malignant conversion when they are subsequently exposed repeatedly to genoioxic 
carcinogens, including PAHs (Hennings e< al. 1983) One ol several reasonable explanations lor the 
consisleni GST-delicient ptienolype in neoplasms that have progressed in White Suckers is ttial multiple 
subsequent "hils" by PAHs or olher carcinogens may have been involved in Ihe later stages ol 
carcinogenesis. Because all advanced tiepalic neoplasms had markedly reduced GST resistance. It Is 
plausible that loss ol GST expression in rare initialed cells woukJ render ihem more prone to further DNA 
damage by agents such as BaP lor which normal hepatocyte GSTs are protective 

This hypothetical concept thai a reduaion in normal cellular resistance lo carcinogens could 
favour tumour progression is imporlanl Irom an epiOemiotogical viewpoint Our findings suggesi thai 
environmental mutagenic chemicals such as PAHs, which can be normally detoxilied by GSTs in these 
fish, are perhaps more likely play a role in the later stages (malignant progression) of carwer development 
in cells tliai have tost GST -dependent resistance mechanisms. This hypothesis implies that tong-term 
exposure to PAHs may necessary to c^ise neoplasms in tissues that are initially rasistant to them This 
view also implies that some species or individuals with genetic deficiencies in specific protedive 
mechanisms would be more susceptible lo carcinogens thai otherwise would be deloxHied. Further 
invBSIigation of this hypothesis and Its Implicaiions (or human suscepibiliiy to the carcinogenic effects of 
environmenial PAHs are currency undenvay The availability ol lish exposed continuously lo PAHs under 
natural conditions provides a means ot understanding the circumstances under which PAHs may be 
carcinogenic to humans This knowledge is essential lo a sound assessment of cancer nek in humans 
exposed to environmental PAHs in contamin^ed water, air or diet. 



B3umannPC3ndHarst±iarg8rJC. (1985). Frequencies oHiver neoplasia In a feral frsh population and 
associated carcinogens. Mar Environ Res 17:324-327. 

DunnBP, Black JJ and Iwlaccubbin A, 11987). ^P-posllatreling analysis Of aromatic DNA adducts in lish 
from polluted areas. Cancer Res 6543-6548 

Farber E and Sarma DSR (1986). Hepatocarcinogenesis- a dynamic cellular perpeclwe. LaD Inveslio 56.4- 

HatMgWH, Pabst MJand JacobyWB. (1974) Gluiaitiione S -transferase The firsl slep in mercapturic acid 
lormation. J B«l Chem 249 7130-7139. 

Harlow HE and Hodson PV, (1988). Chemical contamination ol Hamilton Harbour: A review Canad Tech 
Rept Fish Aquat Sci 1603. 

Hayes MA, Smith IR, Crane TL. HushmoreTH. Thom C. Kocal TE and Ferguson HW (1987) 
Pathogenesis of skin and hepatic neoplasms in white suckats (Catostomus commersoni) from polluted 
areas in Lake Ontario Symposium on Chemical Contaminants and Fish Tumors, Ontario Ministry ol the 
Environment. Toronto, 19B7 

Hendricks JO, Meyers TR, Shellon DW, CasteeUL and Bailey GS. (1985] Hepatocatcinogemcity ol 
benzo[alpyrene to ramlMw Irout by dietary exposure and intraperitoneal injection J Natl Cancer Instil 

Hennings H. Shores R, Wenk M, Spangler EF, Tarone R and Yuspa SH (1983). Malignant conversion ot 
mmjse skin tumors Is Increased by tumor Initiators and unaffected by tumor promoters. Nature 304; 67-69. 

Metcalfe C, Cairns VR and Fuzsmons J. (1987). Distribution o( neoplasms among feral tish in W Lake 
Ontario and eitperimenta! irxJucton of liver tumors in trout win sediment extracts from Hamilton Hartwur, 
Symposium on Chemical Contaminants and Fish Tumors, Ontario Minisiry ol the Environment Toronto 

Meyers MS. Rtiodes LD and McCain 88 (1987) Pathologtc anatomy and patterns ot occurrence of 
hepatic neoplasms, putative preneoplastic lesions, and other idiopathic hepatic conditions In English sole 
(Parophrys vetulus] from Puget Sound, Washington J Natl Cancer Inslit 78333-363. 

Slaga Tj, Fischer SM. Nelson K. and Gleason GL (19801. Studies on tne mechanisms ol skin lumor 
promotion. Evidence for several stages Ol pnjmotion. Proc Natl Acad Sci USA 77 3659-3663. 

Smith IR and Ferguson HW (1985) The assesment of a point source discharge of suspected mutagenic 
and carcinogenic contaminants: an epidemiokigical approach Proc Tech Transfer Conl. MOE 6.285-332 

Sonstegard R and Leathefland JF (1984) Comparative epidemiology the use of fishes in assessing 
carcinogenic contaminants. In: ConlaminanI Effects on Fisheries. Cairns V et al (eds) Adv Environ Sci 
Technol 16:223-232 

Varanasi U, Stem JE, Nishimolo M, Heichorl WL and Collier TK (1987) Chemical carcinogenesis in feral 
fish: uptake, actlvaiion and detoxication of organic xenobiotics Environ Health Perspect. 71:155-170 


nereretica Polluled rolluiod (6 weeks)* 

140 4 

250 B 

120 4 

Vahins ari) aipmisod ai nmoln* ol B.iP sqiiivalnnWq bil« 
'W»iN« siir.tinis h«ld m claan warn lanhs br 6 wasks after captur* 
ttom poHulad art a. 

T able 2 coNCEN r nAT ION or n.ip Mr rAnoi 1 1 rs in nii r or wm i c sucKtns as 


Metabolic Derivative 

Relerence sites Polluled sites Polki led/ fie fere nee 

Elliyl acetate eitlraclable nictaboliles 20 2 

Sullales 6 

Ghjcu'onidos 14,7 

Other 22 6 

20 7 


23 4 


1 2 
16 2 



V.ilui?; Bipiart^ed as nmolas ol EJaP/g bil* 


^M BaP-lrealed 

Polluled site Ftolerence site 

0.59 1.08 

Polluled site 

Vnlusf eipiBSSsd as Wmg ptolatn. 







400 n 










Fipufe \ ; 

Conceniration of B(a)P in the bile of WTiile sucAers from polluted and 
relerence areas at various times after oral dosing with B(a)P (2mg/kgJ. 

Values determined by tola! radioactivity in NaOH extracts of bile ( 1 00 ^ll) 
expressed as nmol/g of B(a)P equivalents per gram of bile (mean ± SE) 








--] ■ 










«... . 

IMIb Iv«i gul gui kidney mU5clB oil I 

„ _ conleni 

Flgur« 3 

ConceniratiDRs of waiersoluBlo 8aP motabolMs (NaOH axOaclabIa} in tissuas and (lu«Js of potlulsd 
and fBlorenco White suckars 2d hours altet orar »xposui» to liSalsd BaP. 

Q polluted 
■ reference 

ta*iey muscle 


Concwntralions ol organ c-soIuBIb BaP melobolites and pafnrn compound (n-H«iana Bxtraciabto) 

to lissues and (tukts ol polluted and relerenc* Wiita suckers 24 hours a(l«f oral B^posuro to ITitialod BaP. 


Figure 4 Radioactivity of Chromatogfaphic Fractions 

ot Bile trom White Suckers from Polluted Sites 

■ Fish1 
n Fish 2 



S e 7 S 5 •■> M 12 IJ M 21 li -ii i* 25 

Retention time (min) 

Figure 5 


Radioactivity of Chromatographic Fractions 
ol Bile trom White Suckers from Reference Sites 

■ Fish 1 
□ Fish 2 


B e ' 9 10 11 1^ 13 14 21 22 i^ '* 25 

Retention time (mln) 



Praliaioarr Data on Plant Bioassaya for t^e Doteotioo 
of BDviroaBent.aL Mutagens io an Aquatio bvironMent. 

Nilliaa F. (irant 

I*epartmetit of Biolpgyj. York University and 

D epart ment of Plant Science, J^. !l._Bo}c_46pS, 

MacdonaM College of McG i 1 1 University. 

Ste. An ne de Bellevu e. faHjebe c H^X^ ICO 


Various higher plant assays have been developed for 
screening and monitoring airborne and aqueous mutagens (de 
Serres 1978; Grant 1S78, Grant et ol. isei ; Nilan 1878). 
These assays are inexpensive, easy to handle and applicable 
to indoor as well as outdoor detection of environmental 
mutagens. Quantitative plant assays for the genotoxic 
detection of aqueous pollutants are relevant and useful in 
establishing water quality standards. This project was 
initiated in order to have a d if f erent relatively 
inexpensive mutagen assay system (using higher plants) which 
would provide a measurement of mutagenic toxicity in assay 
systems not being carried out by the Ontario Ministry of the 
Environment. We are developing such an assay for genotoxic 
aqueous pollutants using two higher plants, namely, 
Tradescantia, and Vicia faba, Kach plant assay has 
different features for detecting gone mutations and/or 
chromosome aberrations. Tradescantia has two assay systems: 
(I) the Stamen Hair Assay for the detection of gene 
mutations, and (2i the Micronucleus Test for the detection 
of chromosomal aberrations . The Vici_a faba assay system 
detects chromosomal aberrations in root tips. These assays 
are being tested to valididate the assays under field 
conditions and to complement information provided by other 
mutagen assay systems being carried out by the Ontario 
Ministry of the Environment. This paper will give the 
preliminary results of a field triai carried out on effluent 
from a paper mill on Lake Superior, a test of the assays at 
Go Home Bay, Georgian Bay, and in a pond at York University. 
Using these assays, we are also collecting data on the 
rautagenioity of a number of dyes used in the paper industry. 



Higher plants provide valuable assay systems for 
screening and monitoring environmental chemicals, both 
gaseous and liquid (Grant at al. 1981). Although higher 
plant assays for the detection of mutagens have been in 
existence for many years, they have only recently received 
the recognition which these sensitive and reliable systems 
warrant (de Serres 1978. Grant 1978; Milan 1978), As Stlch 
and Son (1980) stated "The reoent successful introduction of 
the use of Tradesc ant iB stamina 1 hai rs to detect ai rborne 
mutagens and carcinogens, may be the beginning of the 
recognition of various plant assays which are inexpensive, 
easy to handle and applicable to indoor as well as outdoor 
detection of environmental mutagens". Studies have shown 
that, for n specific chemical agent, comparable results in 
terinc of genetic abnormalities are obtained in plant and 
animal systems. For example, in a survey of studies on the 
effects of pesticides, it has been shown that an excel lent, 
correlation exists between the frequency of both chromosomal 
abnormalities and C-mitoses in plant and animal systems 
(Grant 1978). A similar conclusion was drawn in studies on 
the effect of eight chemicals in several systems, incl uding 
in vitro and in vivo mammalian systems, bacteria, Drosophila 
and plant systems. The plant systems showed excellent 
correlations with the mammalian systems (Clive and Spector 
1976), These higher plant assays are being tested under 
field conditions and were initiated to complement 
information provided by other mutagen assay systems being 
carried out by the Ontario Ministry of the Environment, 
This paper will give the preliminary results of the stamen 
hair mutation assays for field trials carried out on 
effluent from a paper mill on Lake Superior, a test of the 
assays at Go Home Bay. Georgian Bay. and in a pond at York 
University. Data on the mutagenicity of a number of dyes 
used in the paper industry will be presented elsewhere. The 
goal of the study has been to test these assays under 
practical field conditions. 

Materials and Hetbods 

Study SjL^jps 

(at A field trip was carried out to Go Home Bay, Lake 
Superior. July 4 and 5, 1980. Floats as described below 
were placed out at five locations and plants returned to 
York University for analyses. 

(bi A field trip was carried out between July IB arid July 
26, 1 968, to the north shore of Lake Superior, for testing 
the water of Blackbird Creek. Moberly Bay, and Jackfish Bay, 
These si tes are cant iguous waterways and receive the 
effluent discharged from a paper mill. The study was 
designed to test for mutations and chromosome aberrations 


using T radosoant ia and Vlcia faba by tfrowlnd the plants in 
these waterways. Waterways near the test sites were used as 
a control s itea . 

(c> The Tradescantia stamen assay was also carried out In a 
pond at York University. 


A float for holding both Tradoscantia plant cuttings 
and Vicia gerininating seedlings has been designed for 
aqueous testing in different types of aquatic habitats 
' St il 1 , slow moving and fast running water i . The float la 
350 cm in diameter and consists of a 265 cm circular 
plexiglass disk with a central circular enclosed box 10 cm 
in diameter X 4 cm deep which is air tight, with a 4 X 3.5 
cm ring of styrofoam encircling the disk for buoyancy. A 
circular plexiglass rod, 1 . 25 cm diameter and 45 cm in 
length, may be attached by means of a screw to a holder on 
the bottom center of the box on the disk for stability. The 
plexiglass disk is drilled with holes for holding 30 
Tradescantia cuttings. A 50 X 60 cm plastic pot is attached 
to the plexiglass disk by means of plant ties. This plastic 
pot is for holding the germinated seeds of Vicia faba during 

Higher Plant Systens 

(11 TradeBcan^ia clone 4430, heterozygous for flower 
color, blue dominant to pink recessive, developed at the 
Brookhaven National Laboratory originally for the study of 
the effects of irradiation (Underbrink et al. 1973> 

Assays; (a) The staminal heir assay system will detect 
air borne, soil, and aqueous mutants ( Ichikawa. S. 1978; 
Schairer et al. 19781 

(b) The micronucleus assay will detect 
ohromoEome aberrations (Ma 1981). 

(2) Vlcia faba (broad bean) Assay: 

Root tips for the detection of mitotic chromosome 
aberrations (Ma 1982). 


Protocols have been fully developed for both the 
Tradescantia and Vicia assays. A brief nut 1 ine of these 
assays is given below and in greater detail in Appendix 
Tables 1, 11. and III. 

Results on the Trad escantia Btaaten bair assay will be 
reported here. 


Trades cani; i a 

The Tradescantia Stamen Hair Assay is highly sensitive 
to chemical mutagens. In the Stamon Hair Assay, 
inflorescences of the appropriate age are placed in a drop 
of paraffin oil and observed under a dissecting microacope. 
Pink mutant cells are scored. Details of the protocol are 
given in Appendix Table I. 

For the Tradescantia Micronucleus Assay, pollen mother 
cells which are highly synchronized are examined in the 
quartet (totradl stage (about 24 hours prior to anthesisi 
and the number of micronuclej are scored. The micronuclei 
reflect chromosome aberrations which occurred at earlier 
stages In meiosis. The frequency of micronuclei per 100 
cells can be used as an index of mutation. Details of the 
Protocol are given in Appendix Table II. 

Viais faba 

Vicia faba seed are germinated in petri dishes in an 
incubator at 22 C. Seeds are treated and root tips of young 
seedlings are examined for chromosome aberrations. The 
protocol for the Vicia faba Chromosome Aberration Assay is 
given in Appendix Table HI. 

The floats holding the Tradescantia cuttings and the 
Vicia faba root tips were left in the test sites for 24 
hours after which the cuttings weri removed and placed in 
beakers containing Hoagland's nutrient solution and the root 
tips were fixed in 3:1 (3 parts 95X ethane 1 : 1 part glacial 
acetic acid) The solution containing the cuttings was 
changed daily until the cuttings were placed in a growth 
chamber emd data subsequently taken as per protocol. 

Etesults to date 

Trade.?cantiji. §toimen_tiflir Wgt ati qn Asaay 

(a) Go Home Pqiy. Georgian Bay: The five locations were as 
fol lows: 

1. Below the falls, 

2 . I n 1 et , 

3. Open water behind island 

4. Open cove 

t. Cove off channel 

Table 1 gives the results of the Stamen Hair Assays, 
The results are similar with a mean of 3.01 + 0.28. The 
sample from the Cove is higher than all the others; perhaps 
this may be explained by being closer to the shore. All of 
the results are higher than our Laboratory water control 
which averages around I. 01. 


(b) Lake Superior 

Table 3 gives prelinlnary data on mutations from the 
field trial in Moberly Bay and Jackfish Bay (Lake Superior). 
The sites (1 to 6) are in increasing distance froo the 
mouth of Blackbird Creek where the mill waste effluent 
empties into Moberly Bay. The water temperatures during the 
treatment period ranged from 15 to IS-^C and the air 
temperatures from 17 to Zl-'C. The pH was determined of 
collected samples and ranged from 5 97 to 6.34. 

Results of water samples taken from the sites will be 
presented at the Technology Transfer Conference, 

(c) Stong Pond, York University 

Two locations were selected designated the upper and 
lower pond locations. The plants were divided into two 
experiments, in light and dark in which the "dark plants' 
were grown in the dark for 48 hours prior to use. This was 
to determine the effect of travelling with plants in a van 
for 48 hours prior to the plants being placed in a growth 
chamber for mutation analyses. The results from the "dark" 
treatment had « greater range in contrast to the "light" 
treatment, but the results from both treatments were higher 
in the lower pond. 

Conclusi on 

The purpose of the the present experiments was to test 
the higher plant assays under actual field conditions. The 
data analyzed to date indicate that the Tr5d?scaiitia stamen 
hair assay is reliable and can be used to complement 
mutagenic/toxicity assay systems being carried out by the 
Ontario Ministry of the Environment. Data on the 
fflicronuclei assay, Yicia faba chromosome assay and water 
samples are still being analyzed and will be presented at 
the Technology Transfer Conference, 

^kpo wl e dgero ents 

The writer is grateful to Dr. D. M. Logan, Department 
of Biology. York University, in whose Laboratory the 
analyses are being carried out and for his advice and help 
with the project. The assistance of Dr. Michael F. Salamone 
in the execution of the study and his continued interest and 
advice is of immense benefit. The interst and advice of Dr. 
D, Rokosh and the technical assistance of Helen Lee 
throughout this study have been gratefully appreciated. The 
study was made possible through an Ontario Ministry of the 
Environment grant which is gratefully acknowledged, 

labli 1. Tr«diicantia Staaan Hair Hutatton Aiiay ~ 
6d Ko»t Bay, Storgian Bay I«>t Siti* 

Qay BvlDH FalU 

Op»n Hatar Open Cdv* Covt 

Ho. Nu. 
ilautrt pink Floxtri Pinm T]mtrt Pinks Flo«ri Plnlii Flwtr* Pinkt 
•cortd avinti 

4 10 10 




10 r 




































































































Total S9 30) 


66 663 43 2B6 49 





I. 11 





SI. 00 









a) ■ n«an pinka/itaaent b' - titan hai rt/it tatng r) ■ Ptnir autatton*/ 100 

Lab itandard ■ l.Ol + 0.62 autatiani/lOO hatn. 


TABLE I. Protocol tor the Tridfontu SfMn Hair tSTi ftaajy 

The Tradnontu Staaen Hair Assav "is diveloptd b* fl. H. Sparro* 
at Brookhavcn National Laboratorr for trie study of ttit effect* of 
laniiing radiation. Early radt abi ologi tal data deaonstratcd that staaan 
ftain Here sensitive to as litC]e at 0.25 rad of i-ray*. 
Stlch and San M^EO) have itated 'The recent introduction of the use of 
Tradeacant I a staainal hairs to detect airborne nutageni and carcinogen! 
•av be the Deqmninq at the recognition of various plant assays which 
are ineipensive. eat* to handle and applicable to indoor as mil as 
outdoor detect ion of environaental autagen*. * 

For ■nviroriBental cheaicals, the Tradetcantia clone 4430 his been 
developed at (he Broolihaven National LaBorator,. Ihis clone is 
nctcro^ygouf fo' flOHtr color and stdainal hairs. nutations are 
detected by the change in color froa blue to pin^.. 

The Btaainal Hair Bioassy involvasi 

ai nafcing cuttings of young in( iorescsnces prior la aeiosis. 

GroM cuttinqs in aerated Hoagland's nutrient solution. 

bl Expose tie young inflorescences by placing the cuttings 
in the test chenica! and alioHing the liquid to absorb 
through the stea to the inUorcscence. One flower aill 
develop about 40ij staatn hairs. 

d Collect flowers dau / f-oa S to 20 da^s after treataent 

li* ctieaical causes severe tome effect flower developaent aiy be 
delayed, then ei^aeine as long as 25-30 days after treataentl. 
Put flowers in a container with eoistened sponge in refrigerator 

until reaOf tD eiaaine. 

HI ^reo*re slides o, placing > ttaaen m liquid 

oarstiti fsr eiCDsiropic eiaainaticnj add cover slip, 
Evaaine inoeuiately ,i pgesible, or place in 
refrigerator for i»Bt tjav e^aeination. 

e! Ctolojical £:.Baination! Enaained at a aegni f ication of 

<00l no X ocular and a 40 ) oojeclivti. Use white Ugnt 
source (no filters* (or the detection of pink autations. 

f> Stdtistics: E:!aaine and score as in the following emaple: 

Score 18 ttaaens per treataent per day froa IB different cuttings. 
Score daily as above. nutations are recorded as pink events per 
130 or ;00 Etaaens. 

fl coapletB description of the ttchnique ttitb illustrations are gtvan 
in the paper by Schairer et al. (1978i and Underbnnk et al . I1V7S), 
References are given at the end of the report. 



fflftLE 111. Protocol <or the Vicn iabt fl«««v in Eovironaental 
Horn tor 1 na 

Vicn ftttt hat long b«*n used for cvtological and rddiobiological 

studies ifiead 19591. Thii species possess si > pairs of chroao«o**s 
Hhicn are detignated accorditig to centroaere position ai either H 
<*ediani or S 'teriinal). Tht single pair of M cnroBosoatt is aore than 
tBice ttt« length of the S chroHosoaas laean 2.3il) and poiiesses a larga 
satsUita on tfie snort ara. 

ProcBdure i 

In g«n?rat seeds of 'J_. Jaba are fairly large and do not lend 
trisaseWet to qernmate m a petri dish. VaMcus techniques nave Ueen 
developed *or their cjerainatioo and culture. 

1. Seraination i Seeds aay be soal^ed tor 0-12 hours in tap water, triar 
allotted to gerainata in aoist Perlite or Weraiculite or OetHaen paper 
tOHtls, or cotton, or in running theraally regtilatsa tap Mater (Grant at 
ai. I'9l) fo' * days at TO C. Hhefi the priaary roots are batHaan 3 and 
5 ca-long, the seed coats should be reao.'ed and the shoot tip cut off. 
Also, the tips ot the priaar) roots should be cut off to stiaulate 
groHth of secondary lateral roots. 

2, Chaaical a Test liijuids should be prepareo fresn, or tt 
appropriate, are stored in a refrigerator and brouqtit to rooa 
teaperalure c*. 1 hour before use. Tast liouids are changed every day 
le^acorated aaounts aay be replaced . 

Since cheaistry o* coapounfls vary in aode a* action (act at 
different stages u* the cell cycle, S-phaie or during aitosisl, duration 
o' the attotic cycle plus period of ONit tmthesis should be deteramed 
le.g. 20, [8.e, 21 h aitotic cycle). 

'■ Treatwi ts ! For traataents, the feeds are placed on the top of tast 

tubal, or on screens over beakers (6rant et ai. I9eii, containing the 
test solutions so that the roots are subaarged. Ireataent tiaes froa 2 
' 2* hours are aost coaaon with a series ot 2, 4, 8, 12, 2* hours used 
to detaraine tonicity and threshold levels. 

*• Taiparature i The esperiaents shouitf be perforaefl at a relatively 
constant teaperature ca. 20 C for the duration of the enpenaent. The 
enpariaent should be carried out in the Osrl. for the treataenl period. 

3. Ft nation I Z parts 951 ethsnoll 1 part glacial acetic acid, 
prepared freshly for each filiation. fi« 30 mn to I* hours, then aaih 
and transfer to 70'( ethanal for storage in a refrigerator. (Fmation 
and staining Hill suffer after two aonths of storage). 


b. StJintng i Iha Feulgen procedure is Specific iar DNA. rhs roots are 
TBioved (roB the tmatiue and lint trjniferrtd to distilled wdter. 
fitter ;-5 •inutes in water, roots are hvOrolized, 

al Hydrolue in 1 N HCl at 60 C (or 5 am (4 - 11 iin varying mth 
tonditionsi after storage of tissue in aUotiol. the hydrolvsiB tme ii*^ 
need ta be ex tendedi . 

bl Stain in Feulger reagent *or 2 hours 'I -Z hoursl in tht Carfc, 

7. Haceration ; Treat Mith 5', peclmasB for 1-1 hours 'i* left in 
pectinaae too long, root tips will Oecoaie sof t and difficult to handle, 
but 1* not long enougn the cells mil not spread!. 

B. Slide Preparit ion ; On » slia* reaove darkly stained ner ist eiat ic 
region fron the rest ot the root and squash in a drop of 451 acetic 
acid. Mount mth a ;averslip and eaL-e a teeporary seal aith paraffin, 
clear nail polish, ar rubber ceaent. 

'• Perianent S lides i Tenporary sIjiJbs mil deteriorate after a ten 
days. To nate persanent place slide on dry ice end «Hen preparation is 
frojer. coverslip is popped off -ith a ra;or nlade or scapel under one 
of Its corners. Duickly lanerse slide ir 2 changes of aDsoUte alcohol 
and aount with euparol or other nourting nediun and mount with i clean 
covers) ip. 

Hacroicppic Fa raneters i 

11. Turaescence - laroness of root tip; i( high tomctty, roots will 
Hie; pre!:ainar. tests necessary to choose for eKperieent. 

2i . Chanijes of calor - root tips as well as whole plant aay change 
color froa -reataent with certain salts, 
e.g. tluB-green froa copper sulphate, 
e.g. brownish due to towic affects causing cell death. 

3l . Root Forei al C-tuacur 'i-S daysl 

bl bending ot roots ar root tips. 

■51 Foot Length : Coitipare witn controls, 

Hicroscopic Farajieters i 

P. Mitcli; InDe^: Kuaber of dlvidins cells 'all stages of aitoslsJ per 
l^OC abser veg cells, 

2» . Aberrations 0* various types - 

Scor 1 n j i 

Mi totic irden . IC"L' cell counts: t analvsee per series, if Hi too loa, 
discard and go ta new slide with good Ml for scoring. 

Fdf Ly^olagicai Effgcts : cs. ^yu cells scored per lOy cells per root 

Statist ics : fippropnate statistics to be carried out. 

A coaplete description of the technique is given in the capers fay 
brant et ai . ([''Bli. fta (19021 ana rihlman and flndersson 11984). 




Al Melkic and 'fvea Rollln 

Integrated Explorations 

199 MoCurdy Rd., Box I385 

Guelph, Ontario, NIH 6N8 


The fresh water- clan Elllptlo complanata has been used by the 
Ontario Ministry of the Environment for the past 9 years as a 
standard in- situ, bloaccuTULilatinB agent to detect trace 
contaminants in water. The continued use of this popular technique 
has made it necessary to address a number of questions regarding 
the environmental factors which may limit its practical 

To Investigate the environmental factors affecting olam 
bioaccumulation, we tested procedural variables of live olam 
transportation, deployment, tissue processing, and temperature. 
These tests were conducted slniul taneoualy , during a single 21 day 
in-3itu exposure experiment in the Niagara River at Niagara on the 
Lake, The uptake of PCB'3 in E^ eomplanata was used to evaluate the 
effect of these environmental variables. 

The transport experiments involved maintaining clams at ambient and 
ice temperatures in both water and taoist air. Deployment 
experiments Included suspended, flat and compact cages, as well as 
support rings and sand boxes. Tissue prooessing testa included 
holding clams at ambient and ice temperatures for 8 hours and 21 
hours before shucking and freezing. 

Temperature experiments were conducted simultaneously with the 
In-situ tests. Water from the in-situ test area was continuously 
pumped to an adjacent building where its temperature was adjusted 
to provide a range of 5 to 25 degrees celolus at 5 degree 
intervals. This provided olams in the temperature experiment with a 
continuous supply of water from the same source as that of the 
clams being tested in the river. 



Field Methods 


E. cotnplanate specimens, measuring 6.5 to 7.2 cm, were oollectad 

rrom Balsam Lake, Roaedale Ontario and transported to the 

eieper-l mental alte at NlaBara-on-the-Lake within 5 hours, on October 

6tfi, 1986. Details regarding the collection site, methods of 

tranapart, and the test procedures were documented in an earlier 

proceedings which reported preliminary results (Creese et al., 

1986) . 


Clams were transported In contaminant free, food grade plastic bags 
In lake water maintained at the source tenperature of 12 C and 
also at aoblent temperatures which ranged between 15 - 20 "c. Also 
kept at these Cempeature regiemes were seta of clams that were kept 
moist but were not immersed in water. All clams except those 
aoolimated for the temperature experiment were held for US hours at 
12 C. Thermally aocHmated clams were stepped up or down in 5 *^C 
increments every 12 hours until the desired tenperature was 
a t tained . 


ftr environmentally controlled system was established Indoors in 
which 5 aquariums were maintained at 5, 10, 15, 20 S 25 °C 
receiving a constant flow of fresh, thermally adjusted water from 
the Niagara River, This was acoompliahed by chilling water in a 
reservoir to t C and warming water in another to 26 °C then 
connecting them to a manifold system which combined flows from ths 
resevolrs in various proportions to obtain the desired temperature 
Increments. A flow of H l/min. was maintained through the 12 1. 
aquaria which were insulated with 2 inch thick atyrofoam. In each 
acjuarium 6 thermallv acclimated and an equal number of non 
acclimated claraa were supported upright in plastic rings measuring 
2 inches In diameter and 2.5 inches high. The ring rested upon a 
wire meah work to avoid sediment from being trapped within the 
rings. Temperature was monitored every 2 to 3 days for a 21 day 


Clams were deloyed on October Bth, 1986, on top of an underwater 
platform at approximately 3 meters depth. Five experimental 
deployment methods were tested) boxes containing sand from Balsam 
Lake, floating cages suspended at mid depth, ootnpaot meah cages 
maintaining clams jammed together, a standard flat cage allowing 
clams to lay on their side, and ring supports to maintin clams in 
an upright position. The latter deployment method was selected as 
our standard deployment treatment for olams undergoing transport 
and processing testa. Six clams were placed In each containment 
apparatus and these were maintain in the Niagara River for 21 days. 



After a three week exposure perIo{i all clams were retrieved and 
shucked using clean hexane rinsed atalnleas steel knives. Clam 
tissues were drained of excess fluid wrapped In hexane-r insed 
alumlnuHi foil and frozen on dry ice. Tissues were kept frozen at 
-30 C until analyzed. During this tine several processsing 
exprlments were undertaken before tissues were frozen. Live clams 
were held on Ice for 2, 8, and ?H hours before shucking and others 
were held at ambient temperatures for 6 hours. A further group was 
also shucked and kept on Ice for B hours before freezing, fill 
Others were shucked within US minutes of thetr retrieval. 

Laboratory Analysis 

ClaiB tissue analysis of PCB's was done aeeordlng to Ontario 
Ministry of the Environment (1983) protocol with a few 

Clam tissues were thawed, weighed and placed in 50 ml screw top 
(teflon lined) centrifuge tube with 20 ml of concentrated HCl and 
aggitated for 1.5 - 2 hours. Extraction was done with a 20 ml 
portion of 25* dtchloromethane In hexane (v/v5 and aggitated for 
1.5 hours. Approicioately 0.25 ml of 2-propanol was added to the 
centrifuge tubes to break the emulsion layer Chen centrifuged for 
20 ninutea 9 2000 rpm. The extraction procedure was repeated 
twice. Sodium bicarbonate followed by sodium thiosulphate was 
added to the extracts to neutral lie and dry samples respectively. 
Extracts were diluted to 100 ml with hexane and an aliquot 
representing t g of tissue used for cleanup with Florisil 100-200 
aesh (dry pack). The remainder of the extract was used for 
gravimetric lipid determination. Final PCB extracts wore reduced 
to 1 ml using a rotary evaporator, made to 3 ml with 
2,2, U-trlmenthlypentane (iso-octane) and submitted for analysis. 
To increase sensitivity ewtraets of the temperature experiments 
were reduced to dryness before being submitted for analysis. 

All PCB analyses were performed by gas chromatography using 
electron capture detection (CC/ECD). A Hewlett Packard 5880A Series 
GC, equipped with a 30m x 0.32 m I.D. SPB-35 capillary column 
(Supelco, Inc) was used. Typieal GC conditions were: 30 C 
isothermal for 0.50 minutes; then 5^C/mInuto to 300°C, held at 
300 C for 10 minutes. The splitlegs injection system (1,00 minute 
valve time) was maintained at 200 C. 

Levels of PCB's in the samples wore determined by comparison to 
area response factors obtained for standard solutions of Aroclor 
12&0 and Aroclor 125", Injected separately. 

PCB recovery from controls was 99.8 t ♦ 13-3 (mean of n=l2 * S.D.), 
Method detection limit was 8.9 ppb. No corrections were made for 
recovery efficiencies 


TraoBportation Expartaent 

Clania transported in a nolst state (ftTM & ITM) contained 
marginally higher levels of PCB's than those transported in a wet 
state {flTW i LTW) although the levels were not algnlCloantly 
greater ( = 0>05). See Table 1. 

Generally it would seem that the raethod of tranaporting clans to 
the monitoring site Is not of slgnlfloant Importance. 

Table 1. Mean PCB concentrations and standard deviations of 
elams in Transport experiment, n s 3. 
ATM : ambient temperature moist, STW = ambient 
temperature wet, ITM = Ice temperature moist, 
LTW s lake temperature net, SR = support rings. 


in (g) 

0.57 ♦ 0.051 5.93 ± 0.15 

1.1 + 0.053 6.56 + 0. 1« 

0.81 + 0.0U1 6.32 7 0.60 

0.87 + com 5.85 + 0.21 

Deployaent ExperlBent 

Clams deployed in sand boxes had significantly higher levels of 
PCB's than those deployed by other methods ( = 0.051. Levels were 
approximately three times higher than clans in standard cages 
(Table 2.) which are presently In common use. Sinee PCB's are 
normal ly adsorbed to particles, bivalves have to actively siphon to 
aooufflulate PCB's (RIsobrough et al. 1976). This may Indicate that 
the clams are siphoning water more actively, due to 3 more natural 
orientation in the sand box. 

TABLE 2. Mean PCB concentrations and standard deviations of 
olams In Deployment experiment, n = 3- 
LTM =lake temperature wet, SR =3upport ring, 
CC scoDpact cage, FC sfloatlng cage, ^C sstandard 
cage, SB =9snd box. 





III ♦ 26 
31 + 1« 
21 + 10 










21 ♦ 10 

0.87 + 0,0m 

5.85 + 0.21 


27 + 9.5 

0.71 +0.12 

5.2« + 0.22 


5« * 12 

0.7U + 0.031 

6.01 + 0.56 


62 + 22 

0.78 + 0.017 

5.01 + 0.30 


16« * 31.1 

0.21 + 0.030 

6,06 + 0.18 


Procaaslng BxitsriBent 

No algrlfUant dlTference ( = 0.05) «as observed in PCB levels In 
the five processing treatments tested {Table 3.1 indlaatlng that it 
may not be crUlcal ir field logistics do not permit Imniedlate 
prooesslnn of olans. 

Table 3. Mean PCS oonoentrat Ions and standard deviations of 
clams In the Processing experiment, n s 3. 
LIH2 = live ice hold for two hours, LIH8 = live lee 
hold for eight hours, LIHaH : live Ice hold for SU 
hours, SIHfl E shucked Ice hold for eight hours, LAH8 
= live ambient hold for eight hours, JJH = no hold. 









«3 ♦ 17 

0.92 t 0.25 

5.15 * 0.60 

27 + 15 

0.61 t .023 

5.41 ♦ 0.15 


29 * 2 


0.78 t 0.28 

5.10 t 0.29 


33 t 3 


1 .2 t 0.082 

5.7« . 0.U5 


31 t 2 


1 .0 t 0.082 

1.85 t 0.11 


0.87 t O.Olil 

5.85 t 0.21 

Teaperature Experiment 

Temperature dl-l not have a aignifloant f = 0.051 affect on PCB 
accumulation on either acclimated fTable U.) or non-aoc 1 Itnated 
Claras (Table 5.). Aoolimated clana however had PCB levels two to 
four times higher than non-a-iel Ima t ed clans. Only at 20*C was »his 
dtr fere nee significant ( s 0.05) however. 

Table U. Moan PCB concentrations and standard deviations of 
aoelimatad clams In the Temperature experiment. 

n I 3. 









t 25 

* 32 

* 13 
+ 102 
t 3 



0.51 * 0.020 
0.62 t 0.032 
0.5" t 0. 17 
O.itl * 0.12 
1.00 * 0. 1J| 

5.00 * 0.050 
5.31 t 0.19 
5.25 + 0.32 * 0.15 

1.52 7 0.3« 


Table 5. Mean PCB coooentrations and standard deviations 

of non-atjollmated clams In Tooperature experiment. 
n = 3. 

l-£) im) in L£l 


111 + 19 




3U ♦ in 


63 * 15 


103 + 12 

1 .00 + 0.030 

5.18 + 0.26 

0.97 + 0.098 

5.71 + 0.113 

T . 1 1 * 0.05" 

6.16 + 0.26 

1 .01 * 0.070 

U.93 * 0.50 

1 .78 + 0.303 

ll.9« * 0.11 


1> Clams can be transported in the field under more practical 
conditions than has been previously assumed. For example clams may 
be kept moist rather than subflierged In water and be subjected to 
ambient temperature fluctuations of 15 - 20 °C without affeotlng 
PCB bloaocunulation rates. 

2) Creator latitude can also be taken in the field with respect to 
holding clams before freezing. Keeping them on loe for 21 hours, 
shucking and holding for 8 hours, and holding at ambient 
temperatures for 9 hours before freezing result in the same level 
of PCB accumulation as clams that are processed within 15 minutes 
of recovery . 

3) The level of PCB's In indigenous clams may not be directly 
comparable to levels accumulated in clams introduced to a site in 
cages. Clams deployed In sand boxes accumulated PCB's to a level 
three times greater than any of the other ciao cage experiments 
including the standard flat cage. 

4) Varying cage configurations are not likely to be a source of 
error when comparing data from different studies. All experimental 
mesh cages, (flatj compact; floating), as well as the support rings 
used In this study produced statistically Identical results with 
respect to PCB's bioaccumulation . 

5) The seasonal range of monitoring programs may be expanded. 
Temperature within the 5-25 C range did not have a significant 
affect on PCB accumulation on either acoiiraated or non-acclimated 
clama , 

6) Acclimation procedures may actually stress clans and cause their 
lipid levels to be reduced. Acclimated claams accumulated PCB 
levels 2 to 4 times higher than non acclimated clans. This was 
significant at 20 C. 

7) The dynamic interactions Involving PCB bloaoouBula t ion , lipid 
iBetabolisui and physiological rasponsea of claias to stress need t< 
be studied in greater detail in order to improve the resolution i 
clan bioaccumulatlon monitoring techniques. 


Creeae, E., D. Lawls 4 A. Melklo. 1986. Toward the development of a 
standard clam b 1 omonl t orlng methodology: preliminary results. In 
Proceedings of Technology Transfer Conference NO. 7, pp. 205-2T6 
December B 4 9, 1986, Toronto, Ontario. 

Kauss, P.B. & T.S. Saady. 1985. Biological monitoring of 
organochlorlne contatnlnantg In the St. Clair and Detroit Sivers 
using Introduced clams, Elllptlo oomplan atus. J. Great Lakes Bes 

Ontario Ministry of the EnvironBant. 1983. Handbook of analytical 
methods for environmental samples. Laboratory Services and Applied 
research Branch. December, 1983, Toronto, Ontario. 

Biaebpough, R.W., Da Lappa, 8.W. t T.T. Sobaidt. 19T6. 

Bloaccumulation factors of ahlorlnate<l hydrocarbons between mussels 
and seauater. Mar. Pollut. Bull. 7(12):225-228. 

Steel, R.O.D^^aiU J.T. Torrla. 1980. Principals and procedures of 

statistics 2 edition. McGraw-Hill Book Company, Toronto. 633 p. 

Stlokel, L. 1973- Pesticide residues in birds and mammals. In 
Environmental Pollution By Pesticides, ed. C.A. Edwards, po 
254-312. New York: Plenum Press. 




Frank, A.P.C. Gobas, Ronald, H. Russelt and G.O. Haffner 
The Great Lakes Institute 

University of Windsor 
Windsor, Ontario, K9B 3P4 

Introductlft n 

One of the most important issues presently addressed in aquatic 
toxicology is the interpretation of aqueous concentrations of 
toxic organic chemicals in terms of exposure and effects to 
aquatic 1 i fe. Di rect measurement of quantities of trace organic 
chemicals in the water is not only difficult but it is also 
insufficient to determine exposure and effects to aquatic 
organism. Since many organic chemicals were observed to 
accumulate in aquatic organisms, resulting in concentrations in 
the organism exceeding those in the water by orders of nagnitude, 
it was suggested to use organisms to monitor chemical 
concentrations in the water and to determine chemical exposure to 
aquatic life. Organisms used for this purpose such as mussels and 
various species of fish are generally referred to as biononitors. 
The chemical concentration in the bioaonitor is therefore viewed 
as a measure of the chemical concentration in the water and the 
exposure of the organism to this chemical . 6ut in order to 
determine chemical con cent rat ions in water from the body burden 


of the biononitor the relationship between the chemical 
concentrations in the water and the organism has to be 
established. This relationship reflects the organism's ability to 
absorb chemical from the water, its food and other sources and to 
depurate the chemical. In this paper we will review the mechanism 
of chemical uptake and depuration of organic chemicals in aquatic 
organisms and the kinetics of chemical uptake and elimination 
will be discussed. Finally, we will present chemical dependent 
and organism specific relationships relating chemical 
concentrations in aquatic organisms to chemical concentrations in 
the water and show how these relationships should be used to set 
deployaent schemes for biononitors and to interpret biomonitoring 
data . 

UPtakg. depuration and bio accumulation in aquatic organisas 

The expression describing the simultaneous uptake of chemical 
from food and water in aquatic organisms as well as the 
depuration of that chemical to the water (via the gills), into 
the faeces and by metabolic transformation an be expressed as 

dCf/dt = kj.Cw - k2.CF * kA-Cfl - k^.Cp - kR.Cp (1) 

where C is concentration (mol/m^), t is time (h), and the 
subscripts W refer to water, A to food, E to faeces, and F to the 
whole organism (Gobas et al. 1988, 1989) . The organism is 
defined as the whole organism excluding the gjll compartment and 


the gastro-intestinal (GI) tract. kj, kz. 1<a and V^ are 
respectively the rate constants {h"^) of chemical uptake from the 
water, elimination to the water, uptake from food. and 
elimination by egestion in the faeces. kg is the rate constant 
(h-1) for metabolic transformation of the chemical in the 

Foilowing the fugacity approach, discussed at great detail by 
Mackay and coworkers {Hackay and Paterson 1982), equation 1 can 
also be wri tten as 

VF.Zp.dfF/dt = DF.(fw - fp) ♦ Dft.fA - Df-fp - DR.fp (2) 

where V is volume (m-'), Z is the chemical's fugacity capacity 
(mol/m^.Pa) in a phase, and f is the chemical's fugacity (Pa). 
Dp is the overall transport parameter (mol/Pa.h) for chemical 
transfer between water and fish across the respiratory surface 
(e.g. gills). DJ^ is the transport parameter for chemical uptake 
from food into the organism across the gastro-intestinal 
(Gl)-tract. The transport parameter D^ (mol/Pa.h) describes 
chemical elimination in the faeces. Dr (mol/Pa.h) is the 
transformation parameter for metabolic transformation of chemical 
in the organism. The transport parameters Dp, D^, and Og include 
all transport processes involved in solute transfer between the 
water, food, and faeces, respectively, and the solute's final 
storage site in the fish. 
Integration of equation 1 with a constant Cy and C^, an initial 


attempting {but not necessarily achieving) to reach a 
thermodynamic equilibrium. This themodynami c equilibrium is 
characterized by equal fugacities of the chemical in the 
organism, the water and the food consumed by the organism. The 
strength of the kinetic descriptions is that the rate constants 
can be measured directly from uptake and depuration enperiments . 
The fugacity-equations, however, distinguish between 
therraodynamically controlled partitioning phenomena, 
characterized by the fugacity capacity values (i.e. Z) and pure 
transport phenomena, described by transport parameters (i.e. D) . 
Fugacity expressions therefore often give an in-depth view of the 
actual mechanism of the bioaccumul at ion process. The two 
approaches complement each other, and are best combined. This can 
be easily achieved by comparing equations 3 and 4, from which it 
fol 1 ows that 

"l = 


k2 = 


kA " 


kE = 


kR = 



Equations 3 and 4 show that at infinite exposure time an 
organism-water bioaccumulation factor. Kg can be defined for an 
organism simultaneously exposed to contaminated water and food as 

Kb = Cf/Ch = {ZF/Zn).{[t)F/(DF*DE*DR)l * t (f a/^w) ■ (E>a/ (Of + ^e+Db)] } 


bioconcentration factor only reflects organi sm-nater partitioning 
when Df is snail compared to Dp. 

It thus follows that in order to make reliable predictions about 
the bioaccumulation potential of hydrophobic chemicals in aquatic 
organisms and the rate at which bi oaccunul at ion is achieved in 
the organisms, knowledge is required about the processes 
controlling the exchange of solute beti*een fish, water, food, and 
faeces . 

Equation 3 illustrates that when a contaminated organism is 
introduced in clean, uncontaminated water (Cy is zero) and 
consuees uncontaminated food {Ca is zero), it will lose chemicals 
to the water resulting in a drop of Cp with time. The 
differential equation describing this process is again equation 
I, but with a Cw and C^ of lero, i.e. 

dCp/dt = -{k2 * ^E * kR)-CF 


which after integration with an initial CF,fO becomes 

Cf - CF,fO-fe«P(-(k2 + kE + kR).t)} 

In Cp = In CF,t = " ^^2 * ^Z * ''b' ■' 


Equation 16 demonstrates that in a logarithmic plot In Cp 
decreases linearly with time. The slope of this plot is the total 
depuration rate constant (k2 + kg ♦ kf,) and has units of 


Ctiewicol and orqanisp specific relationships for the uptake and 
depuration of organic chewicals in aquatic orqanisus 

Lipid-water mass transfer models were derived by Gobas and Hackay 
(1987) and Mack ay and Hughes (1984) to gain further insight into 
the processes control I ing the exchange of chemical between 
aquatic organisms and water and to develop practical procedures 
to estimate the bioconcentration kinetics of chenicals in fish. 
The authors used the fugacity approach to deri ve the model 
equations but presented their final model in terms of rate 
constants . 

The main feature of this model is that it vi ews the exchange of 
solute chemical between the water and the organism to take place 
in a series of aqueous and lipid layers. All transport processes 
in water phases are therefore grouped together in one overall 
water phase transport parameter Ojj. This overall water phase 
transport parameter contains all transport parameters Dy i in 
water phases. The transport parameters Oy j can refer to 
diffusion, in which case Dy , equals k.A.Zy, where k is the mass 
transfer coefficient (m/s), A is area of diffusion and Zy is the 
cheoicat's fugacity capacity in the water phase. It can also 
refer to non-diffusive transport, where the solute is conveyed by 
a fluid flow G {m^/%) such that Dy j equals G.Zij. The overall 
transport parameter Dy can therefore also be expressed as Qy.Zy, 
where the transport parameter Qy (m-^/s) combines all k.A and flow 
rates G in water phases of the organism. The transport parameter 


When the lipid -water partition coefficient Zt./ZH or Kl '^ 

replaced by the l-octanol -water partition coefficient Zg/Zy or 

Kpy (thus assuming Z[_ to be equal to Zq) equations 20 and 21 

l/k2 = Vl.{(Koh/Qw2h) + 1/Ql} 


1/ki = Vl.{i/Qw * (i/Ql.Kow)}/Lf 


The ratios V^/Ql and ^i/Qv can be viewed as the times of chemical 
transport in Vl m' of respectively lipids and water. However, if 
transport of a given amount of chemical requires a volume V of 
lipid, it will require a much larger volume i.e. Kgw-^L ^^ water, 
since the chemical concentration in the water is a factor of Kgy 
lower than in the lipids. The time for the water phase in the 
organism to transport a certain amount of chemical is therefore 
Kfly times longer than that for the lipid phase. The transport 
time of the water phase is therefore multiplied with Kgy ' n 
equation 22 and alternatively the lipid transport time fs divided 
by Kqw in equation 23. Since the lipid and water transport 
processes occur in series these times are additive and the longer 
time "controls" the bi oconcert rat i on kinetics, 

The expressions 22 and 23 contain two types of variables namely, 
{ i ) biol ogical parameters i.e. Vl, Qy, Ql, G; and Lf, which are 
specific to a particular fish and its physiological condition and 


1/kE - (VF.LF/fio.l-G)-{(6o-LG/QHF)-'(owMGo.Ls/QLF)*l} (28) 

1/Efo " (Go.lg/Qwf)-'^oh * Go-lg''Olf * 1 (") 

where Gj Is th« volumetric feeding rate {in m^ food per hour), Gq 
is the volumetric egestion rate (in m^ faeces per hour), Q^f is 
the water phase transport parameter for cuetnical exchange betnteen 
the Gl-tracl and the final storage site in the organism {in in^ 
per hour), Q^F '* the lipid phase transport parameter for 
chemical exchange between the Gl-tract anil the final storage site 
in the organism {in m^ pg^ hour) and Lg is the organic or "lipid" 
fraction of the gastro-intestin«l contents (in grams of organic 
natter per gram of Gl contents). 

The rec 1 procal of kft, i.e., l/'(J^, can be vi ewed as the t ime 
needed to transport chemical from the fcod into the fish or as 
the total resistance for chemical transfer from the food into the 
fish. Likewise. I/k^ is the time required to eliminate chemical 
from the fish into the faeces or the total resistance for 
chemical elimination to the faeces. The ratios 
{VF/Gi).{Go.Le/QH).Kow and ( Vp/G i ) - (Gq - I-g/Ql) =«" ">• viewed as 
the solute's relative transport times in the water and lipid 
phases, respectively, of the fish or as the relative resistances 
that the solute encounters in the water and lipid phases of the 
fish on its route from the food phase m the Gl-tract to the 
final storage site in the body lipid of the fish. When the 
solute's Kqk increases, and aqueous solubility thjs decreases, 
the water phase of the fish can accommodate only a lower 


concentration of solute molecules. As a result, the time required 
to transport a certain amount of solute with this lower 
concentration increases. The resistance of the fish's water 
phase toward mass transfer thus increases, whereas it reaains 
approximately constant in the lipid phase. For high Kqk 
chemicals, this implies that the uptake rate from food and 
elimination rate to the faeces and thus k^, Efo and kg decrease 
with increasing Kqjj. For low Kq^ chemicals, uptake from food and 
elimination by excretion to the faeces is predoninantly 
controlled by transport in lipid phases, and k^, EfQ, and k^ are, 
therefore, expected to be approximately constant with respect to 

Equation 29 demonstrates that by experimental measurement of Efg 
for a series of chemicals with varying Kqh under controlled 
conditions, i.e., a constant feeding rate and no uptake of 
chemical from the water, it is thus possible to determine the 
fundamental kinetic parameters Qy and Ql- Knowledge of these 
parameters is invaluable for reliable estimation of organic 
chemical bioaccumu lation from contaminated food. 

Gobas et al. (1988) showed that experimental data for dietary 
uptake of chemicals in fish fit this simple relationship with 
values for {Gq- Lg/Qw) . Kqh o^ 5.3 (+/- 1.5).i0-8 and for 
(Gq-'-G^Ql * 1) "f 2.3 ( + /- 0.3), thus resulting in the following 
relationship for EfQ, kyy and k^ 

1/Efo = 5.3.10-8 + 2.3 

1/kA = (Vf/Gj).(5.3.]0-8 + 2.3) 




perforB this "calibration" procedure the rate constants can be 
derived with the expressions discussed earlier as we will now 
demonstrate with an illustrative example. For this purpose we 
will uie a 5 gram fathead minnow {Vp is 0.005 L)with a lipid 
content of 6 %, as a bioironitor. The biomonitor will be deployed 
in a cage to monitor chemical exposure from the water. He will 
assume that the fish is contaminant free at the time of 
deployment. To derive the rate constants for chemical uptake from 
and elimination to the water i.e. k] and k^ equations 22 to 25 
can be used. Equations 24 and 25 show that the Qy and Ql ^°'' *^* 
fathead minnow are respectively 1.4.50-^ i.e. 3.7 L/d and 
0.014.50-6 i.e. 0.037 L/d. Substitution of Qy and Ql in equations 
22 and 23 then results in 

1/ki = 0.00136 + 0.136/Kow 
l/k2 = 8.1. 10-5. Kqjj » 0.0081 



Equations 33 and 34 demonstrate that for tr i chl orobenzene 

(TCB) with a log Kqh of 4.0, kj is 728 d"! and k2 is 1.2 i'^ . For 

mirex witri a log Kqw of 7.5 these rate constants are respectively 

735 d"' and 0.0004 d'' . 

The rate constants for chemical uptake from food and elimination 

to the faeces i.e. k^ and kg can be derived from equations 31 and 

32. Assuming that the caged fathead minnow feeds at a feedinj 

rate of I ^s of its own body weight per day, ky^ for TCB is 

0.0043 d-1 and for mirex is 0.0025 d"'. Assuming that the faecal 

egestion rate Gq is one-third of the feeding Gj and Lp and Lg are 


the water, The relationship between the concentration in the 
organism and the water can now be established by substituting the 
calculated (or measured) values for k ] , k.2 . kf, ^A and 1(r in 
equati on 11 i.e. 

Kb = Cp/Cw = {728/(1. 2*0. 0014+0)} + 

{0.05.10". (0.0043/(1. 2+0. 0014*0))} = 607 


It thus follows from equation 35 that Cy equals the measured 
concentration in the biomonitor divided by 607 i.e. 0^/607. 
However, it should be noted that this procedure is only valid 
when the organism and water are at steady-state at the time of 
sampling. In practice, this steady state will be practically 
reached after 3.0/(k2 + kg + Rr) i.e. 2.5 days assuming a 
chemical concentration which does not vary in time. The fish 
should thus be employed for at least 3 days before the simple 
correlation of 0^/607 can be used to derive the chemical 
concentration in the water. 

For miren the situation is again quite different. With values for 
kj and kA of respectively 735 d'^ and 0.0025 d'^ it follows that 
the chemical concentration in the food of the organism has to be 
approximately 300,000 fold higher than the concentration in the 
water before food and water are equally important exposure routes 
for the fish. However, for a chemical with a Kqw °^ 32,000,000 
simple partitioning of the chemical in the food source of the 
organism may cause such a large difference in chemical 
concentrations in the water and food. Assuming equilibrium 


Equation 36 ttius shows that C^ af^ be calculated as Cf/9.2.10^. 
The tine required to reach this steady state is condition is 

3.0/{k2 + •<£■•■ '^fi) i-^- ^^^^ days. This shows that for mirex the 
biononi tor has to be deployed for a much longer time than when 
TC8 is being monitored . I t may even be possible that the 
blomonitor will never reach this steady state condition within 
its 11 fe time. The body burden for mirex in the biomoni tor Is 
thus a function of the deployment time t. This time function is 
given by equation 3, which after substitution of the values of 
the rate constants is 

Cp - {735. Cw/(0. 0004 ♦ 0.00083 + 0)} + 

{0. 05. lO'-S.Ck,. (0.0025/(0.0004 * 0.00083 * 0))} (37) 

The example discussed above shows the necessity to interpret 
blononitoring data on a chemical spect fie basis. It can thus be 
concluded that when biomoni tors are to be used as a tool to 
neasure chemical concentrations in the water and chemical 
exposure the kinetics of chemical uptake and depuration have to 
established for each chemical of interest. 


We gratefully acknowledge the Ontario Ministry of the Environment 
for financial support. 


Norstrom, R.J., A.E. HcKinnon and A.S.W. deFreitas. 1976. A 
bioenergetics based model for pollutant accumulation by fUh. 
Simulation of PCB and methyl mercury residue level in Ottawa 
River yellow perch (Perca flavescens). 0. Fish Res. Board Can. 28 
: 815-819. 




Zsoll E. Kovaii and Jan J.H. Ciborow&ki 

Depariment of Biological Sciences 

University of Windsor 

Windsor, Oniario. N9B 3P4 

Onlario Miniiiry of the Environment 

Technology Transfer Conference 

Toronto, Onlario 

28-29 November. 1988 


Benihic aqualic instcl larvae living ia contaminated sedimcnt& accumulate ^significant 
organoihlorine burdens, However, ihcir value as indicators is limited by sampling diffi- 
culties and the necessity of acquiring enougb biomass for analysis. The nocturnal, 
pholophilic, winged adull stages are more easily collected than larvae. Our objectives 
>*ere to assess seasonal variation in adult insect availability and coniaminant burden, 
contrast concentrations in animals from contaminated sites with those from unconiami- 
DUled areas, and determine minimum sample biomass that provides reasonable detection 
limits for organocblorine contaminants (PCBs, pesticides and others). Light trap col- 
lections yielded large samples of Trichopiera (mostly Cheumalo ga.jj h si from late May to 
late August at sites on the Detroit and St, Clair rivers. Ephemeropicra t Hexa g cnJa . 
Cacnis l were abundant for only 1-2 weeks in midsummer. Midsummer collections yielded 
many Trichoptcra at five sites along the Niagara Riser. Howcicr, low temperatures 
limited the size of catches of Trichopiera, Hexapenia and ^ .'spn jij at four locations on 
the St. Marys River. Contaminant conceniraliuns in animals from Detroit River samples 
were typically 1-2 orders of magnitude greater than in animals from several central 
Onlario control sites. Series of triplicate subsamples of different mass from a single 
large collection were analysed for 30 organocblorine compounds. The proportion of con- 
taminants at delectable concentrations and median coefHcient of variation stabilized in 
samples >0.4 and >(),K g dry mass for animals from contaminated sites and uncnntaminatcd 
sites, respectively. Sisasonal variation in contaminant concentration at sites on the 
Detroit and St. Clair rivers will be compared with spatial variation among major Great 
Lakes connecting channels. 



Organochlorioe coniaminaliixi of ihc Lduicniian (ircai Litkes has caused increasing 
conccTQ in the pasl iwa itt;cadi;<i li i\ now generally accepted [hat sediment contamina- 
tjon ma> influence ihc eniite fauna o( a walct hody ihrough upiakc b\ henlhic inverlc- 
braics and subscquenl transfer ihtough ihc (nod chain lO higher irophie levels (Slruger 
et al. I9SS). Con»quently. biological monitoriog of scdimcnl contaminants ha^ been the 
focus of much research in recent years. 

Degree of contaminBoi bioaccumulaiion by aquaiic animals depends on the type of 
exposure and the chemical ptopcriica of the compounds (Reynnldson l<t87), Bcnthic inver- 
tebrates live on or ttithin the sedimcnl^i aod are exposed to organic eoniatninanis ihrough 
direct contact and feeding. Freshwater mus^cK (Kaus^ and Hamdy l'385), Chironomidae 
(Diptera) (Larsson 1984), oligochscte worms (Olivier l'.'K4). and caddisdy latiac (Bush ct 
al. 1985} have all been reported lo carry high organochlorine contaminant burdens 
Since contaminant hody burdens are often proportional to concentrations in ihc sediment 
(Larsson 1984), benlhic invcrictirales arc potcniially useful as indicators of sediment 
con lamination- However, ditficuliies encountered u hen sampling boitom-duelling organ- 
isms and the need For extensive processing often limit the amounl of tissue available 
for soalyiet. 

Collection of the adults ol henihic insects presents a cost-cf fective alternative 
to hentbic sampling. Caddisfties [Trichoptera) and mayftic (Ephemeropiera) spend most 
of their life as larvae, uitbin or in contact with the sediments. The nigbl-active 
winged adults emerge during the summer in large numbers. Adults are shortlived, do not 
feed or defecate, and with the exception of a small proportion of contaminants shed with 
the larval skin (Larsson 1'>H4). body burdens remain unchang.-d following emergence. 

Mauck and OUiin (l')77). Clements and Knwatski (lQ!t4), and Ciborowski and Cnrkum 
(19H8) analysed adults of aquatic insects collected near Urge rivers and detected 
elevated levels of organothlorine contaminants, indicative of sediment concentrations. 
Though these studies demonstraled the potential for use ol these animals as indicators 
of sediment contamination, there is a need for studies evaluating insect seasonal avail- 
ability and dispersal, spatial variation io contamiodni bjtden. and minimum useful 
sample size for conlaminanl analyses. Our previous work (Kovats et al. 1987) has shovkn 
that adult aquatic insects can be collected in large numbers using light traps, and are 
available in sufficient numbers for analysis throughout the summer Preliminary results 
of dispersal studies have shown that active dispersal b) adults is limited. 

Id this paper we compare conlaminanl concentrations in adult aquatic insects col- 
tecied at coniaminated and unconiaminated sites. We also evaluate analytical precision 
associated with different sample weights and the effect of differences in storage tem- 
perature and length of storage time on analytical results. Such data are necessary if 
standard techniques for routine monitoring are to be developed. 


2.1 Sample Collect ion and Preparation 

Aduli aquaiic insect sample> collected from mid-Mav to mid-September 1987 were 
analyzed (or 30 urganochlorine contaminants (19 PCB congeners. S pesticides. ocUchloro- 
slytenc <OCS). hexachlorohcn/ene (HCB). peniachlorobenfene (QCB)) by gas chromatography 
(GC). Insects were collected with modified Pennsylvania lype light naps (Frost 1957) 
placed at 4 sites along Ihc Detroit and St. Cljir rivers (Table I) and at three sites in 


Tahte 1. LocalioQh of sampling staEions.. 

Latiiude LoBgilnde 

Mb Riifil Designation (North) West) 

I Detroit River Canard 42''l r48" SSOOe'lS" 

3 DcUoii East Windsor 42''30"27' 82°56'56" 

i S( Clair Sombra 42»4202- 8«2903" 

-i St Clair Sarpi. 42°S4'12- S2^2r29- 

central Ontario. Detroit and St. Clair River ledimeDts contain elevated levcU of 
organochlorine coniamindots and the rivers have been designated Areas of Concern (Inter- 
□ attonal Joint Commission. 1985). Central Ontario sites were chosen to represent uncon- 
laminated areas. Sampling was also conducted along ibe St, Marys and Niagara rivers in 
I9S8 to evaluate the applicability oF our methods accross broader geographic regions 
(Kovats ei al. In prep.). 

The catchment reservoir of the light traps contained dry ice, which immobilized 
insects without introducing contaminants. Detailed collection procedures and specific 
•.ample dates were outlined hy Kovats et al. (1987). Samples were separated into consti- 
tuent taxa (Hciiagenia (Ephemeroplera). Hydropsychidae (Trichoptera). Other Tricfaoplcra. 
and Other Taia) using hexine-rinsed lorceps and stored at -20 or -TO'C (see below) prior 
to analysis. 

Triplicate 2-5 g samples (fresh weight) of insects were prepared for coniamioant 
analysis. Sample mass used depended on availability. An additional, similar-si/ed 
portion was dried al IllS'^C for 24 h and rcweighed to estimate relative moisture. The 
dryjfresh weight ratio of this sample was used to eslimale dr) weights of GC-analyzed 
samples Replicates from each site were extracted and analysed on different days. One 
solvent blank and 5 insect samples were extracted and analyzed on any one day, 

2.2 Gas Chromatographic Analysis 

Samples were homogeniy.ed with mortar and pestle in 50 g Na^SO^ . Solid-liquid 
extraction was employed io extract contaminants and lipids (20 g Na,,SOi^and 30(1 mL 50''l 
dicfaloromeihane (DCM)-5u^t hexane). The extract was concentrated to 5 mL by rotary 
evaporator and the concentrate was added to a Biobeads column (S-X.1, 200-40U mesh). Two 
fractions were eluted by addition of 301) mL iS'^i DCM-55';"t hcxane mixture. Solvent was 
evapoiaied from the first fraction and the remaining lipid residue was weighed. The 
second fraction containing all extracted organochlorine compounds, was concentrated to 2 
mL and cleaned by passage through a column containing 8 g Florisil. Two tractions were 
eluied by hexane and 50^. DCM-50^ hexane, respectively. Both were concentrated to 2 mL, 
diluted with heiane to 10 mL. and 1-jiL portions were injected into the GC. Specific 
conditions and methodology employed during gas chromatography were as outlined by Cibor- 
owski and Corkum (1Q8K). Contaminant concentrations were quantified based on peak 
patterns by comparison to those on chromatograms of standard mixes of known organochlo- 
rine con cent rations. Recovery efficiencies were evaluated by spiking uncontaminatcd 
samples with known amounts of standard mixes. 


2.3 Sample SlortgeTime and Tempetatute 

The effect of storage lempeniurc wai evaluated by unjiyzing injects collected at ■ 
eonlaminaied Detroit River site OVindsor) and at an uncontamiriBled central Ontario site 
(Gull River). Samples were split and stored at different temperatures (-20 and -TO'C) 
for 4 mo. Portions of I lacgc (281) g) sample (Detroit River, Windsor) Fro/en at -2(I°C 
were analyzed al 60 day intervals to determine the effect of length of storage lime on 
analytical results. Concentrations were compared by one-way analysis of variance 
(ANOVA) (Soksl and Rolf, ]<)69). 

2.4 Minimum Useful Sample Size 

Minimum reliable sample weight was determined using samples of Hcxa i fenia from the 
Detroit River (Windsor, collected on 2J June 1987), and Hvdr,ip5vche (Trichoptera: Hydro- 
pa ychidae) from tbe Gull River (central Ontario, collected on 18 June 1VH7.). Tripli- 
cates of S different lubsample wcighlh of insects (O.Oy, O.IH, 0.38, 0.75 and 1.5 g, dry 
weight) were analyzed from single collections, and coefficicais of variation were calcu- 
lated for the 30 compoundi at each sample weight. Median ocffieicnts of variation were 
piolted againbi sample dr) weight. The weight at which the change in median approached 
icro was considered to be the minimum allowable sample weight yielding acceptable varia- 
tion among triplicates. Percent nortdcteciahle compounds was also plotted against sample 
dry weight. 

2.5 Iniersiie Compsriiont 

We compared contaminant conceniralioni in samples of insects collected at purport- 
edly unconiaminaicd central Ontario sites to those from the Detroit and St. Clair 
rivers. We expected to find significantly higher concentrations of all conidminanis in 
samples from Detroit and St. Clair Ri\er sites. In addition, similar taxa ( Hexagenia 
and Hydropsychidae) were analyzed from different sites alo^g the Detroit and St. Clair 
rivers to assess possible spatial trends in contaminant di.itribution along the rivers. 
To simplify comparisons of organoch lortne conlaminani levels among samples from various 
sites, we selected 4 representative compounds (Table 2). Three of the compounds chosen 
each corresponded to one of 3 major groups of contaminiinls distinguished by Ciborowskl 
and Corkum's (IVSS) principal component analyses of data generated by collection and GC 
analysis oT adult aqualic insects dt sites along the Detroit and St. Clair rivers, to 
addition, the pesticide dieldnn was selected to represent pesticide compounds. Site 
comparisons were performed using nine. way AN OVA. 

3.0 tESDLTS 

3.1 Sensitivity 

Detailed results of contaminant analyses (site comparisons only) are listed in 
Tables 3 and 4. Our analyses detected a mitilmum of 25 of the 3D contaminants studied in 
all samples from unconiaminaicd sites. Recovery efficiencies were >90% in spiked 


Table 2. Repr«SieatalTve ccimpouDds used lo simplify presentation of ihe data set and 
ibe major groups of compounds they repicscnt. 

Represenlativc compound 

Group of fompnunds 

PC8 !80* 
{:.:-.3.4,4',5,5 -Hepiachlorobipheiiyl) 

PCB 66 




Highly chlorinated FCBs 
(heia-, hcpta-. and octa- 

Less highly chlorinated PCBs 

(icira- and pcniachloro- 


Pesticides (Aldrin. 
Dicidiin. Hcpiachlor. 
Hcptachlor epoxide. 
pp'-DDT. pp-DDE, 

* PCB aumberiog follows BalKcbmiier and Zell I9S0. 

3,2 Sample Storage 

Neither :>loragc lempcralurc nut length of storage lime signiricanily influcoced 
dnalylicat results (p>U.U5J. No degradation was noted in Bnimals io either of the 
fractions of saropies split and stored at -20 and -70°C for 4 months, and results of GC 
aaalyjis were identical. Similarly, a difference of dO days in length of storage lime 
had no effect on contaminant analyses. 

3,3 Minimum L'scfu I Sample Size 

Coefficieoi^ of variaiioD calculated during minimum sample siie experiments and 
percent oon-dcieciahle compounds wete plotted for Detroit River Hexagenia and Gull River 

(eeoiral OnUrioi Hydropsvche (Figure 1). Based on lhe*e plots, sample dry weights of 
at least 11.38 g ( Hexagenia . 25 animals) and 0.75 g t Hvdropsvchc . 170 animals) arc 
sufficient to provide acceptable repeatability of analytical results in samples from 
highly cuntaminaied and relatively unconlaminated areas, respectively. 

3.4 Inlersite Comparisons 

Couceniraiifins of the representative contaminants in Hexagenia collected at 3 site* 
along the Detroit and St. Clair rivers were compared to (hose at Balsam Lake (Figure 2). 
Significantly higher levels of contarainaots were detected in samples from the Detroit 
and St. Clair rivers (p<0.05. one way ANOVA). Exceptions were dieldrin at Station .1 
(Sombra. St. Clair River) and PCB 66 at Station 4 (Sarnia. St Clair Rivet), concentra- 
tioos of which were not significantly different from those at Balsam Lake. In both 
cases large variation among replicates reduced power of statistical tests. Significant 
spatial variation in contaminant concentrations In conspccific animals wiihln rivers was 













" ■•" 
















11 '"l 






11 •• 

■ (•••I 

i< In 


m 'm. 




It ••II 



m i'*i 






ii 111 




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■ •■ 



■ *• 







(t i»j| 








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1 w. 





11 u 










• ■# 




tbi. <> 

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11 .111 




11 u 







Tifale V M»n 

1.1 S.E. 

ulioii aI pe^iiciJi: 

!'.. KvilciDSlii: Din 

nd lUhrr orftiaiKhla 
n[ ihe ciinpaiindi i 

■ compiiuh Ji \nt kft'drv wri^hl) m rcpic^cDiiiivc jdull iqujli 
iiMi-d in Appcodn 1 (Htpi - H.'piaehlti', H. Epoi.- Mtpuchlut 

ND - Don .IcIccKhIr) 

|L<)LATI()N LlFimaj QLfl- 









(Sin. 4) 

So* bra 
(Sin. S) 


(Sin. :) 

n Caniiij 



















I in 

(II IW.') 



1. 52 

;7 4B 

( ^ mt>i 


10.2] i;o in 

(0.777) (I.VWi;) 


i.»» :ioi 
fp-OKSi tSLSiiL 








2\ n 


Iti h4 



(0 26.1) 



45 7(1 
(tl 255) 

211 111 



2(1 II 













(0 KOR) 

t 41 


; 27 

2 .W 0.5(1 

(n.sB;i (0..1S1) 

4.72 .1.71 

(1.08.1) (2.06;) 


J- 5. I 


<4. J1M» 

14.47 5 70 

(l.O.W) ((1.1%) 

12 41 6 10 

(I.NNI) lOAM) 

Ih.T! 10.26 

(1. 3761(2.447) 

6 49 12.70 







in 160) 





(1. 114) 

1 77 ;:.4o 

(0.062) (1.252) 

Nr>5 IV R7 

(0.277) (i.6tU) 

B.7,1 12.24 

(0.74«) (4,!J5) 

14 44 ^142 

(0.574) (3.2V41 






1 80 




018 038 

j { i. 




















009 018 038 075 

Dry wl. (g) 





z 20 



018 a38 
Dry wt (g) 









018 038 
Dry wt {gj 



Figure 1. Coerriciem of varialion (above) and percent Dondetccublc rompounds (btlow) ptolled 
againsl sample dry weight fDi (A) Heiag Cpifl al Ibc Detroit River (WJadsor). (regression 
i-qualion: %Nondeieciable=25.33 x (Dry weighl)EXP(-0.430), and (B) Hydropsychidac at Horseshoe 
Dam (central Oalario), (regrcsstoo equation: %Nondetectable=l4 23 x (Dry wcighl)EXP(-0,4SH). 
Median values arc reprcseolcd by large open circles. 








*— ' 























2 3 A 



PCB 66 

















2 3 4 








PCB 180 

2 3 A 



Figure 2. Concenlrations of "ieleclcd coniaminanls in Hu-xagcnia ac ihrec Detroit and St. 
Clair livet stations and ul BaUam Laltc (BL). In^uf fi<:i<:nl mdlctiiil wai. collected 
at SUtioa 1 (River Canaidj (or analysis. Vertical bars represent 1 S.E. (n-3). 


alsa obiervcd. HCB and PCB conccniralions in HeKiyenia decreased fTom Slition 2 (De- 
iron River, upsiream siic) lo Slatioo 4 (St. Clair River, upstream site). This paiicro 
corresponds lo ihai reported tor PCBs ia ihe sediments at [hese sites (Pugsley el al. 

Coalamiaaai coacen I rations were stgniricanlly higher in Hydropsycbidae at Station 1 
(Detroit River) (ban at central Ontario sites (Ausabie River and Gull River, Figure 3), 

Pcslicide concentrations were eievaied in samples from the latter sites when compared to 
Icieli of other contaminants. The higher coneeotraiions most lilicly reflect local agri- 
cultural activity. 

Collcclioas along the Niagara River yielded large numbers of Triihoptera al all 

sites. Sizes of catches from St, Mar>s River sites were limited by low lempetaiutes and 
were considerably smaller. All s»mples were numerically dominated by Tricboptera, with 
Ihe exception of one sample collccied al the mouih of the St. Maryi River, which con- 
laincd large numbers of the mayfly Cacnis (Ephemeropiera: Caenidae) Whereas Si. Marys 
River Trichoptera samples were quite diverse. Hydropsycbidae and Lcploceridae (Tricho- 
piera) dominated Niagara River samples. Contaminant analyses of these samples are in 

*.o ni^icussioN 

4.1 Minimum Sample Si^e 

Results of our analyses indicate that larval aquatic insccis accumulate organochlo- 
rine compounds and retain sufficient amounts as adults lo permit precise analysis by GC. 
Minimum sample i'u.e experiments indicate that sample biomass considered lo be small for 
other ioveriebraie tissues CI,; - ;.5 g, fresh wt.) is adequate 10 provide acceptable 
precision in analytical results- This can be explained by the relatively low water 
conteol and high lipid content of adult aquatic insects. 

■t.2 Spatial Variation 

Comparison.s of contaminated with unconiaminalcd sites yielded results in accordance 
with expectations. Exceptions were the pesticides, which exhibited higher levels than 
cRpeclcd at uncontaminated sites. Although there were still significant differences 
between Detroit River and central Ontario sites, this demonstrates the ubiquitous occur- 
rence of these compounds. Variation in contaminant concentrations in the insects among 
locations paralleled those in the sediments. 

Use of samples of adults lo infer local larval concaminanl concentration requires 
the assumption that animals have emerged in the immediate vicinity of light traps. 
However, both active and passive dispersal by flight can occur. Preliminary results of 
dispersal studies suggest that active dispersal by Ephemeropiera and Trichoptera is 
limited: In July 1987, Lake St, Clair Hcxagenia dispersed mean (il S,E.( distances of 
2570i22B m, and hydropsychid caddisflies travelled 970-178 m (Kovats and Ciborowski. in 
prep). Passive dispersal by wind is also limited, since aquatic insect adults seldom fly 
under windy conditions. During inclement weather they cling to the substrate or lo 
vegetation {Johnson 1969). The pronounced spatial trends in adult contamination de- 
tected by our study also argue for limited dispersal. Mauck and Olson (1977) and 
Clements and Kawatski (1984) also reported significant spatial variation in contaminant 
distribution in Mississippi River Hexj g t inia samples eollected UI-20 km apatl. We have 
investigated Iwo alternative trapping methods that could be minimize inclusion of adults 





^ ^ 
















PCB 66 

^ m 






PCB 180 



Figuic 3. Cuncenlraliuns of selected conlaminanis in Trichoplera from Ihc Detroil Rivet 
and two uncontaminalcd silfi (DR: Deiroii River a[ Windsor, AR: Ausable River, GR: 
Gull River). Vertical bars represent 1 S.E. (n=3). 



Overall, adult aquatic idsccis arc sunsitivc iadjcaiors of aquatic coatamiDaiiun 
and yield reliable data regatdiog the degree scdimeol contan I nation in the area sur- 
rounding a given (ample station. Though thi^ approach docs not provide specific esti- 
mates oF scdimcDl contaminant concentrations, it is useful in assessing contaroinatioo on 
a larger scale. Animals captured during single collections represent the major aquatic 
insect tana of <2-5 km stretches of rivers or laitcs. Besides its utility in providing 
monitoring at single sites on a repeating basis, collcclion and analysis of adult 
insects may be especially well suited for preliminary surveys of areas previously not 
studied, due to its relative simplicity and cost-cf feclive nature. 


We wish lo thank Stephen Peraai for assistance with sample collections and pro- 
cessing. Dr G.D. Haf fner made available gas ch torn aiugra phi c facilities at the (ireat 
Lakes Institute, and Dr R La^ar provided advice on analytical procedures. We thank A. 
Hayion and W, Scbeider (Ontario Ministry of the Environment) foi assistance with various 
,i>pccts of this project. This research was supported by R.A.C. Grant PLSiJ from the 
Ontario Ministry of the Environment, 


Ballschmiicr. A., and M, Zcll. 1980. Analysis of polychloftnated biphenyls (PCB) by capillary gas chromatography, Ftes. Z. Anal. Chem. 302:20-31. 

Bush. B., K.W, Simpson, L, Shane, and R.R, Koblint;. 1985, PCB congener analysis of 
water and caddisfly larvae (Insecta: Trichoptera) in the upper Hudson Rivet by gas 
capillary chromalogiapby. Bull. Environ. Conlam. Toxicol, 34:96-105. 

Ciborowski, J.J.H. and L.D, Corkum. 198S Organic contaminants in adult aquatic insects 
of the StClair and Detroit Riven, Oniatio, Canada. J. Great Lakes Res. I4:14B-156. 

Clements, J,R and J. A. Kawatski 19H4. Occurrence of polychlorinated biphcnyU (PCB's) 
in adult mayflies <]lc»iJiJLttia bilincala) of the upper Mississippi River. J, 
Froshwai Ecology. ;.611-6I4, 

Frost, S.W, 1957. The Pennsylvania insect light trap, J Econ, Entomol, 50;287-292, 

International Joint Commission. 198S, Report (jn Great Lakes wuatcr Ouality I98.S. Great 
Lakes Water Ouality Board. Windsor, Ontario, Canada. 

Johnson, C,G. 1969. Migration and dispersal af insects by flight, London: Mcthucn. 

Kauss, P.B. and Y.S. Hamdy, 1985. Biological nioailoriiig of organocfaloriae coniamioaDts 
in the St. Clair and Detroit Rivers using imroduccd clams, Elliplio complanalus . 
J. Great Lakes Res. 11:247-263, 

Kovats, Z. E,, J J,H. Ciborowski, and S, Pernal, \9H7. Biomoniloring protocols tor adult 
aquatic insects: Collection procedures, seasonal variation and dispersal. Proc. 
1V87 Technot. Transfer Conf., Toronto, Ontario. 



[b*( have dispersed long diiianccs. Picliminaty results of collections using boai- 
mouDtcd traps, which caich emerging msyrty subimagoes, are promising. Wc have also 
employed underwitei light traps to collect benthic larvae, but these did not caich 
appreciable numbers of organisms. 

Comparison of our data to similar aoHlyses of Ciborowski and Cotkum (1986) reveals 
the poieniial for considerable year lo year variation in coniiminant body burdens in the 
taxa studied. Though different extraction procedures were used during our analyses, the 
effect on contaminant concentrations was relatively small (unpublished data). During 
tbe period of one year, concentrations of indivi4ual PCB congeners increased 2-10 fold 
in Tricboptera at Slalioo 1 (Detroit River, downstream), white body burdens of other 
orgaaochlorine compounds remained unchanged. PCB concentrations in Hetagenia also 
increased slightly at Station 2 (Detroit River, upstream), and at Station 3 (St, Clair 
River, downstream) but dropped by approximately 50^' at Station 4 (St. Clair River, 
upstream). Based on these findings, downstream transport of PCBs is a possibility, 
though inclusion in upper St. Clair River samples of HeKayenia that may have emerged 
from lest contaminated southern Lake Huron areas may account for the relatively low 
contaminant burdens found in animals at Station 4. Moreover, since both our present 
study and that of Ciborowski and Corkum (198S) represent single collections from each 
site, it is possible that the observed temporal variation represents time-specific 
sampling error. Nevertheless, wc believe that the changes in insect contamination 
reflect temporal trends in sediment contamination. Verification of seasonal trends 
requires reliable sediment data, precise localization of sample sources, and repeated 
sampling to estimate the degree of random variation. Analysis of data collected during 
1988 will provide more conclusive evaluation of seasonal variability. 

4.3 Choice of Taxa and Seasonality 

The animals used in our study were chosen primarily on the ba%is of availability. 
However, microhabilal differences within the Trichoptera also warrant consideration when 
selecting lava for monitoring contaminants since the extent of contaminant uptake and 
the type of compounds accumulated are dependent upon larval feeding behaviour and sub- 
strate preference. Bush et al. (W85) analysed caddisfly larvae of various species from 
the Hudson River and found significant differences in PCB body burdens among members of 
different families. We therefore recommend that samples used for monitoring be ideally 
composed of single species or at least mcmbcri of the same family. The Hydropsychidae, 
composed primarily of riverine species, appear to be ihe most appropriate group for 
maniloring contamination in large rivers, and the mayfly He^agenia seems well suited for 
moniioring lakes. 

Though adult aquatic insects emerge throughout tbe summer months, time of sampling 
for the laxa selected for contaminant analyses <ihould be carefully chosen. Some tricho- 
pteran species are bivoliinc, the second generation emerging in late summer (Mackay 
1978), having developed from eggs laid in early summer. Sampling for these animals 
early in the emergence season ensures that contaminants were accumulated during a stan- 
dard length of time (approx. 1 year), defined by the animal's life cycle. Since He«a- 
gCOil has 1 relatively short emergence period, typically 2-3 weeks, sampling for these 
animals must be precisely timed. 


Larsson, P. 1984. Transport o( PCB't from aquatic to tctreMrial cnvitonmenis by emerg- 
ing chironomids. Environ. Follul. 34:283-289. 

Mackay. RJ 1979 Life history palierns of some species of Hydropsyche (Triehoplera: 
hlydrapsycbidae) in souihern Ontario, Can. J. Zool. 57:963-975, 

Mauck. V-.L and L.E, Olson, 1977. Polychlorinaled bipheoyls in adull mayflies IHexagenia 
t)il in^;a ^ a ' (rom the upper Misti&tippi River. Bull. Environ. Conlam Toxicol. 

Oliver. BG I9S4. Uptake of chloTiDatecI oigaaict from aathropogeDically coolamlnaled 
sediments by oligochaelc uorms. Can. J. Fish. Aquat. Sci. 41:878-883. 

Pugslcy. C.W., P.D.N. Heberl, G.W. Wood, Ci. Broiea and T,W Obaf 1985. Disiribulion of 
cunlamioanii in clams and sediments from the Huron-Eric Corridor. 1 ■ PCB's and 
ociachloroslyrenc. J Great Lakes Res. 11:275-289. 

Reynoldson, T.B, 1987. Inuraciions between sediment contaminants and benihie organisms. 
Hydtobiologii 149:53-66. 

Sokal. R.R., and F,l. Rolf. 1969. Biometry. San Fraocitco: W.H. Ftecman and Co, 

Struger, J., D.V. Wesctob, D,J Hallell, and P. Mineau. 1985. Organocblorinc contami- 
nants in herring gull eggs from the Deiroil and Niagara rivers and Saginaw Bay 
(1978-1982}: Contaminant discfiminaais. J. Great Lakes Res. 11:223-230. 



An Ecosystem Approach to the Monitoring 
of PCBs in Pristine Ontario Lakes 

CD. Metcalfe and C.R. Hacdonald 

Environmental and Resource studies Program 

Trent University, Peterborough, Ontario 


It has been estimated that 7.5 jug to 24 ug of PCBs are 
deposited per square meter per year from the atmosphere into 
lakes in the Great Lakes region {Swackhamer and Armstrong 1986, 
Murphy and Schinsky 1983), and that atmospheric deposition nay 
account for up to 80% of the PCBs entering the Great Lakes 
(Thomann and Di Toro 1983). There is potential, therefore, for 
PCB contamination from atmospheric sources in more northern On- 
tario lakes. Because of their hydrophobic nature, PCBs tend to 
accumulate within lake biota and lake sediments, and because of 
their low rates of degradation, these compounds may remain for 
long periods of time in these environmental compartments. Con- 
centrations of PCBs in some sport fish species have risen to 
moderate levels (0.5 to 1 ppm) in even "pristine" lakes of On- 
tario. It is the objective of this study to survey the levels of 
PCBs in all environmental compartments in seven Ontario lakes to 
assess the extent of atmospheric contamination, and to indicate 
the pathways for movement and loss of these compounds in each 
lake ecosystem. 

Lakes chosen for study (Figure 1) are within the Ontario 
townships of Peterborough (Rice Lake), Durham (Lake Scugog) , 
Stanhope {Boshkung Lake, St. Nora Lake), Oakley (Wood Lake), and 
Sebastopol (Lake Clear), and from Algonquin Park (Lake Opeongo) . 
The selection of these lakes was based upon trophic status, 
degree of lake development, and level of PCB contamination in 
lake biota. Based upon Ontario Ministry of Environment monitor- 
ing data for sport fish, PCB contamination in these lakes ranges 
from low (<10 ppb in smallmouth bass), to medium (10-100 ppb) , to 
high (>100 ppb) levels. Rice Lake and Lake Clear, which have the 
highest levels of contamination, have known point sources of 
PCBs, while the other lakes in this study have no distinct 
sources of these compounds. Three of the study lakes are con- 
sidered moderately eutrophic (Rice Lake, Lake Scugog, Lake 
Clear), while the other lakes are oligotrophic. Among the 
oligotrophic lakes, there is a gradient in terms of lake develop- 
ment, from Boshkung Lake (most developed) to Lake Opeongo (least 
developed) . 

Samples of water, suspended particulates, sediment, 
zooplankton, crayfish, clams, pooled insects, and 5 species of 
fish (golden shiners, bluntnose minnows, YOY yellow perch, adult 
yellow perch, smallmouth bass, lake trout) were collected in 


1986, 1987, and 1988 for analysis of PCBs . Sediment cores were 
collected from the deepest deposition zone of each lake using a 
KB corer, and divided into three sections of 0-3, 3-6, and 6-9 
cm. Sediment extracts were prepared by sonication, and sulfur 
compounds were removed by precipitation with mercury. Lake water 
samples (54 L) were filtered through 0,3 p.m glass fibre filters. 
The filtrate and filters were extracted with methylene chloride 
for analysis of PCBs in water and suspended particulates, respec- 
tively. Zooplankton were collected using a 276 ;a mesh conical 
net, and other biota were collected using dip nets, minnow traps, 
seine nets, trap nets, and by angling. Among fish species col- 
lected, an attempt was made to sample constant body sizes between 
lakes, and the ages of fish were estimated from scale readings or 
published age-length data. Samples of muscle from the mid-dorsal 
region of each fish and from the tail of crayfish were used for 
analysis. Biota samples were extracted into hexane using 
soxhlet apparatus, and lipids were removed by gel permeation 

Sample extracts were cleaned up and subf ract ionated by 
silica gel column chromatography into a "PCB fraction" containing 
PCBs, DDE, aldrin, heptaclor, and lindane, and two other frac- 
tions containing organochlorine pesticides. The PCB fractions 
were analyzed by high resolution gas chromatography using a 
Varian 3500 gas chromatograph with a 30 m DB-5 capillary column 
and EC detector. Rather than analyze all possible PCB congeners, 
19 congeners were selected for analysis, ranging from trich- 
lorobiphenyls to decachlorobiphenyi (Table 1) . These compounds 
include most of the major congeners present in commercial PCB 
mixtures and in environmental samples (Oliver and Niimi 1988, 
Duinker et al 1988, Norstrom et al. in press) . 

Analysis of samples from the study lakes supports the 
original monitoring data used for lake selection. The concentra- 
tions of total PCBs (sum of the 19 congeners) in the muscle of 
adult yellow perch ranged from 8 ng.g in Lake Opeongo to 9L 
ng.g~ in Lake clear. The same gradient in concentration is ob- 
served for all other species of biota in these lakes. In addi- 
tion to the differences in the concentration of total PCBs in 
biota from the seven study lakes, the patterns of PCB congeners 
in the biota differ between lakes (Figure 2). This may reflect 
different sources of PCB contamination and/or variations in the 
ecology of the biota In each lake. 

The levels of total_PCBs in the biota of "pristine" lakes is 
approximately 4-190 ng.g ^. Within lakes there are variations in 
the pattern of PCB congeners in the biota samples (Figure 3) . 
Principle components analysis indicates that biota in the lower 
trophic levels have congener patterns most closely resembling 
Aroclor 1248, while the congener patterns in upper trophic levels 
most closely resemble Aroclor 1260, There are a broad range of 
congeners present in the sediments from these lakes (Figure 3) . 


The data for concentration of PCBs in the various trophic 
levels of pristine lakes, when expressed on a wet weight basis, 
indicate that PCB concentration increases with trophic level 
(Figure 4). These data would seem to support the concept of 
"biomagnification" of PCBs through the food chain. However, when 
PCB concentrations are expressed on a lipid weight basis, there 
is no clear association between trophic level and PCB concentra- 
tion. The accumulation of PCBs in lake biota may be governed by 
a variety of factors, including lipid content, feeding ecology, 
the age of the organism, and the organism's relative metabolic 

Detailed analysis of PCB data has been completed for Lake 
Clear (Macdonald et al, submitted). This lake was accidently 
contaminated with PCBs in the inid-1970's and contains the highest 
concentrations of PCBs among the lakes In this study. Concentra- 
tions of total PCBs in sediments were highest (597 ng.g ) in the 
surface section of cores, which indicates that PCB transport in 
the lake is still a dynamic process. PCBs in lake water and 
suspended sediments were easily detectable at approximately 1.9 
pg.L"^ and 870 ng.g"^ dry weight (1.0 )jq.L~^), respectively. 
Concentrations of PCBs in biota were high (up to 2900 ng.g In 
lake trout muscle) . Unlike the pristine lakes in this study, 
PCBs in Lake Clear biota were uniform in concentration and con- 
gener pattern across all trophic levels. Relationships between 
bioconcentration factors (BCFs) and octanol-water partition coef- 
ficients (log Kq^) of individual congeners indicate that very 
hydrophobic congeners (log K^^ >7) are accumulated in biota to a 
lesser extent than would be predicted from their lipophilicity 
(Figure 5). Several hypotheses might explain this finding, in- 
cluding reduced bioavailability of very hydrophobic congeners due 
to binding with dissolved organic or colloidal material in lake 

The final objectives of this study are to describe the dis- 
tribution of PCBs and other organochlorine compounds in the seven 
study lakes, and to establish criteria by which atmospheric 
deposition of PCBs into lakes can be recognized from point-source 
discharges of these compounds. We will also use environmental 
fate models developed by the USEPA (i.e. WASTOX) to simulate at- 
mospheric inputs and the movement of PCBs throughout the ecosys- 
tems of these lakes. Using these models, we hope to determine 
the baseline contamination levels to be expected among biota in 
pristine Ontario lakes, and to predict the rates of decline (if 
any) in levels of these compounds among all compartments of the 
lake ecosystems. 



Du inker, J.C. , A.H. Knap, K.C. Binkley, G.H. Van Dam, A. Darrel- 
Rew, and .T.J. Hillebrand. 19B8. Method to represent the qualita- 
tive and quantitative characteristics of PCB mixtures: Marine 
mammal tissues and commercia] mixtures as examples. Mar. Poll. 
Bull. 9:74-79. 

Hawker, D.W. and D. . Connell. 19BB. Octanol-water partition coef- 
ficients of polychlorinated biphenyl congeners. Environ. Science 
Technol. 22:382-387. 

Oliver, 8.G. and A.J. Niimi. 1988. Trophodynamics analysis of 
polychlorinated biphenyl congeners and other chlorinated 
hydrocarbons in the Lake Ontario ecosystem . Env iron . Science 
Technol. 22:388-397. 

Hacdonald, C.R. , CD. Metcalfe, T.L. Metcalfe, and G.C. Balch. 
Distribution of PCBs in a small, contaminated lake in Ontario. 
Environ. Science and Technol. (Submitted). 

Hurphy , T. J . and A . W . Schinsky . 198 3 . Net atmospheric inputs of 
PCB's to the ice cover in Lake Huron. J. Great Lakes Res. 9:92- 

Horstrom, R.J., M. Simon, D.C.G. Muir, and R.E. Schweinsburg. or- 
ganochlorine contaminants in arctic marine food chains: Iden- 
tification, geographical distribution, and tempera 1 trends in 
polar bears. Environ. Science. Technol. (In press). 

Swackhamer, D.L, and D.E. Armstrong. 1986. Estimation of the at- 
mospheric and nonatmospheric contributions and losses of polych- 
lorinated bi phenyls for Lake Michigan on the basis of sediment 
records of remote lakes. Environ. Sci . Technol. 20:879-883. 

Thomann, R.V. and D.M, Di Toro. 198 3 . Physico-chemical model of 
toxic substances in the Great Lakes. J. Great Lakes Res. 9:474- 



Figure 1: Location of the seven study lakes in central and east- 
ern Ontario, Canada. 

Figure 2: Relative proportions of the PCB congeners analyzed in 
adult yellow perch from 3 "pristine" study lakes (Wood, Boshkung, 
Scugog) and troin 2 lakes receiving PCBs from point sources 
(Clear, Rice) . 

Figure 3: Relative proportions of the PCB congeners analyzed in 
the sediments and biota from different trophic levels in Boshkunq 

Figure 4: Concentrations (^jg.g"^ wet weight) of the PCB con- 
geners analyzed in the biota from different trophic levels in 
Boshkung Lake. 

Figure 5; The relationship between the bioconcentration factors 
(cone, biota/cone, water) in Lake Clear biota and the octanol- 
water partition coefficients (log K ) for individual PCB con- 


Locotion of Study Lokes 

r^'^ I 




















lit l««a-^i...^ 






t,! lU) IM nn 1^.1 1M IWI IIOIDI in >B9 IH 

Congener Nurnbcr 




lokc trout 

smallmouth boss 

odiiX yellow perch 

X YOY ycil 

YOY yellow porch 



la 11 M n « iDi »j MB HI ne'M iM'Mwowi iwna '8' JO* 
(a, ("1 (».X'«1 

Cong oner NumbOf 



laVe trn 

!-.FnnllrTioiilh bdSS 

ntliill yel'ow pcrcti 

I>lunln05(? mmnow 

VOY /gIIovk peicfi 



I II 5J « »■ >(i. «' MO ir.M-is.> imiBDiwwi >••<•» iM »• 

Congener Number 












1 - 

• crayfish 
' ~ "O golden shiner 

Q zooplonkton 

-■- '£^ smallmoulh boss 

Log K 



TibU 1 - Sunnary of PCO congenen anilysod fn icven iluJy Itkct. nrickets 



'"9 "<«.' 








2. 2", 5 




i.4-,S 12.4.4) 











2. 2'. 3. 5' 












!.3,3'.<'.6 (3,4.3-.4'J 







6. 74 [(i.67J 


[?.!■, 3. 4'. 5', 6| 



6.9! (G.SO) 

(7. 3. 4. 7', 3', 6') 




7.!', 3. 4. 4', 5' 











7, 7". 3. 3', 4.5, S', S- 




7. 7". 3, 3', 4. 4', 5, 6' 








7. 7M. 3', 4. 4'. 5. 5' 



7. 7', 3. J', 4. 4'. 5, S-. 6. 6- 


fUPAC nuMfioring lyilcn for PCS cangencri. 
I09 K„ viluDj froM llaHkcr jixl ConncH |t9S8). 




A. A. Crowder 

W. Dushenko 

J. Gretg 

Queen's University 

DeparcnenC of Biology 

Kingston, Ontario K7L 3S6 


Diversity and bio mass of aqua tic nacrophytes dlmlntahed during 
hypereucrophlcatLon of the Bay of Qulnte, Lake Ontario, during the 1960s and 
have not recovered. Numbers of watsrfowl and manmials In wetlands tn the Bay 
^re also tow. Tt was hypothesized thai the wetlands are contaminated by 
diHtals derived fron Bines in Che Moira Valley, compounding the stress of 
ejcrophlcaClon. Possible contaminants analysed In samples of sediment have 
included Ag, As, Al. Cd. Co. Cl, Cii. Hg. Mg, Mn. Na, Hi. Pb. Ti . V and Zn. 
Nutrient concentrations In sediment were also measured (N, P, K, Ca and Mg), 
El^oenCal concentrations In eoergent and submerged plants and In snails have 
been analysed, using neutron activation analysis and atomic absorption 
apectrophotnmecty, to teat for transfer of contaminants up food chains. 
Cover classes of subaerged plants at sices around the Bay were correlated 
with netals and nutrient concentrations, and also with limiting factors such 
a<, exposure and slope of shores, organic carbonates and silt content of 
sediments, and pH, 

Significant differences occur between wetlands in the Moira River area and 
in Hay Bay, about 20 kta east. Concentration of As, Co, N'a, Mn and Pb are 
higher in Hay Bay. concentrations of As (2A-U.'> ppm). Cr (29.2-46.6 ppm), 
and Cu (10.3-25.1 ppm) are potentially toxic, while sediment Hg is rot (< 
0.2 ppm). Submerged plants in the Moira area contain significantly higher 
concentrations of As and Mn ( means 4.6-6.8 ppm As; 656-703 ppm Mn). One 
sampled snail apecles ( Stagnicola elodcs ) accumulates 34.93 - 12.44 ppm Cu. 
Concentrations of Mn and \l in snails are also elevated, but show high 
variance within species. 



The International Joint Cointlaslon has recognized two areas on the north 
a ho re of Lake Ontario which need remedial action prograpnes , Hani It on 
Harbour and the Bay of Quint e. The latter has historically been the aont 
productive fishery In Lake Ontario. The shore of the Bay has extensive 
wetlands, which were estimated in 1979 as 2824 ha of emergent aarshes and 
627 ha of subaerged and floating-leaved vegetation (Crowder and Brlstow 
1986}i These wetlands affect the aquatic ecosyatea, by providing food 
and/or habitat for Invertebrates, aaphiblans, fish, birds and nassals. The 
wetlands are also used for nuskrat trapping and for non-consunpt Ive activity 
such as blrd-watchtng. (Cf. National Wetlands Working Group 19B8, Ch. 6). 

During the early part of thla century eutrophlcation of the Bay of 
Quinte increaaed, culminating In s hypereutrophlc state In the I960'8, which 
led to control of point sources of phosphorus in the 1970'b (Minns et al. 
1986). The weedbeds of subaerged aacrophytes at first increased and then 
sharply declined In bloaass during the hypereutrophlc phase. Such declines, 
coabined with lowered diversity and Invasion by non-native plants, have been 
characteristic of eutrophlc alcea both in north America and in Europe 
CCrowder and Brlstou 1988). By the 1970'b the area of weedbeds had become 
limited by light penetration (because of dense algae), and diversity and 
blomass were low In coaparlaon with regional noma. In Che emergent cattail 
•Cauda diversity was also Low, and both weedbeds and aarshea were vlalted by 
fewer algratory birds than are expected In such habitats In eastern Ontario 
(Crowder et al. 1986). 

Metal contamination frequently accompanies eutrophicatlon, as a result 
of Industry, urbanization and agricultural run-'off; Hamilton Harbour is an 
obvious case of eutrophicatlon with simultaneous metal contamination of 


sedlBtnts. In ihe Bay of QulnCi? sedlnent In deep water is known to contain 
elevated concentrat lorns of Cd, Hg, Pb and Za (Sly I9B6). An obvious source 
of metals U the Inactive mines of the Mol ra Valley, where contaninatud 
sediments containing Pb, Co. VI, Cu, As, Ag and Hg were analysed by Mudroch 
,ind Cnpoblanco (1979, 1980) nnd by Pealke ec al. (1982). 

In the area near Belleville and Big Bay, subwerged weedbeds during the 
(Jfcsde 1972-82 had less biooiais and cover than In other areas of the Bay ot 
qulnte (Crowder and Brlstow 19S6}. We therefore hypothesised that this area 
<aay have received netala from the Hoira valley over j long parlod of time. 
This project, supported by the Ontario Ministry of the Envlrctnoent and the 
World Wildlife Fund, is dealgned to test this hypoche'ils, and to estimate 
thi? likelihood of transfer of metals In local foodchains. The project will 
ilso test for organic pollutants In wetlands, during 1988-89. 

Three sets i>E data have bfen collected. The first conies from Bend Bay, 
1 riverine wetland below the mining area on Che MotM River, wher^ sediment, 
plants and Bnlnuls could be collected tu represent a 'worst possible case'. 

The second consisted uf A9 sites along the shoreline, J»t places with 
dnd without vegetation. These sites were <in the north and south shores of 
the Bay, which is about 90 km long. This set was designed to give an 
uvBtvlcw of near-shore conditions throughout the Bay and is referred to as 
near-shore samples. 

The th Ird set was collected wlt>\ln foiir wetlands with submergent and 
emergent vegetation. Two wetlands, at Point ftnne, were as closo ss possible 
to the mouth of the Molra River, and were chosen to represent sites with 
potential contamination over a long period of time. The second pair of 
wetlands, aacchtng the first In aspect and vegetation structure, wens In Hay 
Bay, about 20 km downstream from the Hoi ra ai w^r, and therefore presumably 
free from cnnt^mlnat Ion. This set contal ned both near-shofM .ind on-shor.: 



tamples. Sftea have been aapp«d In CrowdtT ec al. 19B6a & b. ReeuUs 
Including analyses ot sediments, plants and (Ullinals have been published tn 
Crowdar dC al. . 1988a. b and c. This report therefore condenses Information 
based on field work done In 1987. Some analyses are not yet complete, and 
will be published with reaitlts of the second phase of research (which «1 1 1 
Include seasonal measurenentii of uptake , f racttonatton (spec! at Ion) of 
metals and analysis of organic cnntamlnarici. Species reported hare Include 
MyriophylluB splcatuw , Eur.^slan al If ol I, whl ch is the *osE coaiMon submerged 
plant Ifi the Bay and ValUsnerta awerlcana . wild celery, a major food for 
ducks fCrowder and Brlsiow 1988). Consuner organ Isns had to be «pecii?s 
found sbiindant ly and not likely to migrate tar, - those reported here are 
the snails Stagnicola elodea and Planorbella trlvolvls , the mofC abundant 
ffloUuscs found In nany sites. Buth species of snail are eaten by fish and 

Although the Bay of CJuJnte hai b^en sudled Intensively {Minns et al, 
1986), the hydrology of Ua shorelinea and wetlanda are not well known. I'. 
is not known whether uleaientB draining fron the Molr-i callings have been in 
particulate f(,riii or In tolutlon. The aiacshes are subject ta wind set-up and 
to sma 1 I seiches, but their Intensity and f rt-quency ari^ iiiiknotfri, .m is thei r 
potent tal for sertiment transport. It ti; iiJt kniiwri whether wat^r carrier 
sediment up Into the raarsheii durhig the melt, or erodes thera; Che compl-.'icl ty 
■)f thi; shoreline presumably could allow both occurences slmijlcaneously, but 
there ouiy be temporal variance In amount and velocity of the meltwater, and 
in tho ice in the marshes. Ceis (1985) has described 'Ice-Eoot' phenomsna 
In the St, Lawrence Ri ver which af feet -led tment -ind plants, and si mi la r 
events were observnd In Way Bay djrl ng 198a. In on -shore si to-j spatial 
dlstributl.n of rii*.i.als has been studied, to try t'l elucidate possible 


deposit ional/eroBlona! paccerna, and ulll be reported later. 


Sediment collection 

At near-shore sites sedtnent was coltacEail vlth an Gknan dredge. In a 
depth of 0.5 ■ of water. At on-shore sices (e.g. Point Anne, Hay Bay) 10 
onposiCe sanptes of 6 cores each were collected, using plastic coring Cubes 
ar shovels. Only the rooting zone, the uppernoat 10 en of -led tnent , uas 
utilized. Field laensureqMnts of pH wer^' oiade. Particle ttie analysis, 
using sieve and hydroaecer techniques, wa* done in the Hydrology Departaent 
>->f Loyalist College it Selleville. Loss of Ignition (organic carbon) was 
lae^s'ited after 2 h at A20° C. Alt sedlaent tras air-dried and sieved to 2 fM 
JIaah!ter before •element analysis. 

toUect ion of plants and measurement of vegetation cover 

At near-shore sites WyriophylluB aplcatua and Valllsnerta americana 
wer-; collected a? cli>"ie is possible to sediment collifctliin points, «her« 
vegetac Ion occurred. In th.; on-shore si tei coaposltu samples of 10 plants 
if each species wera collected at each coring point. Plants wer-j washed and 
j>1ac>?d In 1 cooler tn plastic bags In the Fl^ld, Roots and ahoots werp 
Buparnted for blon-iss efitlfflitlon, alr-drted at 'O' C and ground to ^ 
diameter of t ma prior to analysis. 

Cover was listlnated as ttiur classes, ranging from • iio |)l.intq to * ■ 
dt'nsii cover, in lata suram^r. 
Collection o f snails . 

Thirty-five Planorbella t rivol vis and IS Stagnl cola el odea were 
collected EroB each on-shorn site. Snails were kept In delonlaed water for 
24 h for digestti'e clearance, then frot^n. Tissue was later removed from 
shells, and drtsd at 90° C. Small tamples -Jer« not drouod. 


Eleaent analysis 

i. Meutron activation analysta (NAA) 

Protocol and ^tandardliation of NAA using a Slowpoke-2 reactnr at Royal 
Military College, Klngacon, were developed by Dr. !. Poland (Queen's 
University Analytical Services) and Dr. P. Beel«y. Two allquots of m dry 
HBraple of aedf nent , plant or artlnal I (ssue were wr>l gheA In a 7 mL pi as tic 
vial and heat -sealed. Samples uere I r radiated In an automated handl Ing 
system. One sample received a 1-3 minute irradiation at a neutron fluK of 5 
x lo'' n era*' sec" , followed by a decay tt»e of 1 - 10 m to detect short- 
lived nuclides (e.g. Kl, Ca, CI, Cu, K, Mg, Mn, Na, Tl and V). The second 
aawple was Irradiated for 2 h, and li>ft for a decay period of (a) 100 h to 
detect fts, Br, K, La, Sa, Sb., and Sc and (b) 250 h to detect Ba, Co, Cf, 
Cs, Fe, Hf, Rt>, Ta, Th, U and 7.n. 
ii ^t oil c absorption spectrophotowetry 

Extraction was done by twu method-;: 
(a) 1-2 g of dry material «cre ue[gh<>d Into a teflon beaker and dissolved 
using llNOj, HCIO^ and HF, followed by re-acld 1 f lea t Ion using 202 HCl. 
Staod-ird-i were prepared in 2DS HCl, and Ag, Cd, Co, Cu. Nf, Pb and Zn 
di^frraioi^d by flame AAS. (b) Dlgeicton uUh HNO^ and HjOj was done In 
teflon 'bombs' for 12 h at no**. No significant Jlfference «aa found 
between extractions (a) and (b), therefore the first laechod was used alonn 
for later tuits. Deteral nation of Hg used a c"ld vapour technique and Aa. 
was Measured using a hydride generation system. 

Certified r>-feroiice niacerldls fur ecandarij tx ing analyses were BOSS NRCC 
marliU! sediment, \&S SRM 157 2 cltrun leaves and TORT-1 KRCC Inunrtebrate 
t Issue. 


Statistical analyalfl 

Stat graphics progranoes C references) were used far data handling and 
.■injlyais. Significance was indicated by p values less than 0.05. 

Cover of plants in near-ahnre sites. 

Cover was estimated as 4 classes and was posic Ively correlated with 
sLlt and with organic natter (loss on Ignition) In sediment. It was 
negatively correlated with sand and fetch; fetch la a measure of the 
(■xpaaure of a site to wind and wave action. High energy shores have sand 
disposition and tend to be too disturbed for plant growth. Positive 
i:>jrr«l at ions with silt and organic matter Indicate that at sices where 
iiccunuliitlon can occur, both of fine particles and plant debris, dense 
(irowth of plants Is possible. fCf. Crowder et al. 19fl8b). 

Cover was also related ro pfl, possibly because sites accuaiilailng 
.-ganic debris become slightly more actd. Significant correlations were 
found between cover classes and Pb ini 7.n [n sediment and the nutrients P 
md Mgi these correlations ware positive but there was a negstlv" 
ri^larlonehlp with Cu. Correlation coefficients nee Mhown In Table I. 
IClemgnts in near'shorc aedlwents 

Ranges and aeans of selected eleMencs are given in Tabl« 2. 
-iraltatlona and advant.iges of technique have been discussed In Crowder « 
il_, (19883). The broad range of eleaentt analysed by NAA al lowed an 
overview of the 49 sites to be laade. Some elements whl ch were expected tn 
occur were generally near limits of de tectabi H ly , notably Hg and Cd. 
Absence of these potentially conic element* Indicates a niajor difference 
between the near-shore sedtoents tnd the sites In deep water described by 
Sly (1986). They also differ ■jtrongly from sites up the Moira valley 
described by Mudroch and Capobianco (1979), The range for Cd, less than 1.0 


Cu 2.9 ppBi, Is comparable en surface values In deposit lonal zones In Lake 
Ontario (0.1-6.2 ppm) reported by Hudroch et ai. (1988), Their value for 
Hg In surface sediments in deposit ional basins was 0. 140- 3.9 S ppw, and Che 
mean Qulnte uctliind values were all lesq than 0.1 pprn. 

ChroBluB had a ntean value of 31 ppm; Its range tjf 14-68 ppn Is within 
the llTalts {8-133 ppm) reported by Mudroch et^ al . (1966). Copp«r ranged 
from 24 to 490 ppm, whereas the surface range in deposlcional basins given 
by Mudroch e£ «_1. (1988) is 26 - 109 ppiB. In wetlands Clooschenko (1988) 
has reported high variance, and li agrl cultural loi I a mean of 2^ ppm for 
Cu. The situs with high eopp»?r a'p therefore patent la 1 ly toxic. 

Nickel had a me^n of 20.9 ppm, which Is at the lower value reporced by 
Mudroch et »!.. CI98S); this value is very^ close to those in Rattray Marsh 
and Coate's Parxdlfe reported by Gloo^chenko (3) -29 ppn) and higher than 
the mean Cor agri cli 1 1 ura 1 soil (16 ppm). :!Inc, IHo Cu, had vr.ry high 
varlancf, with a nw^n .'F ^t.J ppm. yudruch et a_L. (ISSS) also show very 
liirge ranges (e.g. 16-1225 ppm in tmbayraencs) while Che Quints w-'flands are 
again very similar t^> the marihe^ described by Clouschunlco (1978). with 29- 
88 ppo. Lead, with a leean of 15 ppn, Is relatively \<rj except at the upp«r 
«nd of the rang!-, 'y'i ppm. Hiidr.ich et^ al^. (1988) a'"'' values oE 7-285 ppm 
for deposit I onal ba-ilns, and 7-l(i9 ppm in river moiiths. 

Sodium ranged f row 0.05-4. 7 percent . a high range for a f r*?shwHter 
systen. Chlorine was also high ,it sone sices but did not coltclde with the 
peaks for Na. Salinity coiit rols plsnc diversity In cii^stal marshes In the 
United Scales, utch ttie lowest species rlchnesi In saline sites (Larrick and 
Chabreck 19 78) , the r-? fore the ()uinte values IndlcsCi possible condic ions of 
low diversity for plants. Sidluni is high In soim? t al lings In the Moira 
valley, but both Na and CI could hr. derived from road salt and snow piles. 


Analysis of As frow the near-shora sites Is not yet cunplete. 

Several dtst rl but itinal patterns were apparent (Fig. 1). Nickel was 
high only at Bend Bay. and In low concentration froo the mouCh of the Hoi ra 
River downsireaia. Copper was highest at Bend Bay but also peaked In Hay Bay 
and Sucker Creek. Zinc was concentrated at several sites, although sho-lng 
■JOB* general attenuation do-nstreaa. Lead seens to have an urban origin, 
belna highest at the mouth of the Moi ra River rather than close to the 
mines. Copper was not closely related to particle size, but Zn, N) and Pb 
wrire p3sictvely correlated with fines, which could explain chelr 
dl-itrib.ition patterns. (Table 3). Lead and Zn, and Cu and Nl tended Co co- 
Elenents in on-shore sites 

The greatest difference from near-shore sites was the higher organic 
content ( 531 mean loss on Ignltlonl; th« m«a-i value for near-shore site-. 

WA» 161. 

Arsenic ^t on-shore «Ues Is ilgnl f Icantly greater In concentration aL 
Point Anne oarshes than downstream In Hay Bay Barshes CT«bU 4). Arsenic 
value-! wer.; strongly and positively correlated wUh organic matter and 
«paUly correlated with Pb and Zn. The .Ignlflcant difference betwen these 
j^i lan<i« Kupports ch,- hypothesis that nine drainage has affected wetlands In 
th^ upper Bay of tjulnte. In coaparUon with Rend Bay. however, the A3 
v-ilues are a magnitude smaller; value* froa Bend Hay, Point Ann^ and Hay Bay 
nvirshes are shown In Table 4. Co and Nl were also higher «C Berid Bay than 
In all the Bay of Quint- marshes. Then; was more Pb aurf ^i near BellevllU 
than in Hay Bay, but Cu w-ia similar in both areas. 
Metals in plants 

Concentrations of As In both Hyriophyllun splcfltuni and Vallisneria 
amerteana were significantly higher In Point Anne than Hay Bay wetlands. 


reflecting sedlmsat values. (Tablo 5) The planLs had specific dirEerences 
in uptake, for exampU Pb was significantly higher in Hyflophyllum and Na in 
Valllsngrla at both areas (Table 6). In both species, and in both ueclandE, 
Hg was less Chan Q.2 pptn. Copper, which was negatively related to cover in 
near-shore yices. had rna^n values in plants between i.O and 5.3 ug. g -I. in 
both areas. 
Metal content of snails . 

Planorbella tflvolvis and Stasnicola elodes accumulated s Lgnlf tcant ly 
different amounts of Ca, Hg, Al. Cu. V, K, and Na in their rissues. P^ 
Erivolvls had higher Ca and Mg, and S^ elodes had high Al and Cui Us mean 
Cu eontenc was 34.9 ug.g" . '\rsi.-nic In snails in the onshore marshes r-jng«d 
from 1.8-4.i ug,g~' in P^ trlvolvis and from 2. 5-12. ^ ug-g" In S^ elodes, 
with the higher value in Hay Bay. Planofbella trlvolvi^ can accumulate 
higher anjoiints whtre con t ami hat Ion Is ^r^^'itct, an r.hc nienn value from Send 
Bay uan M ug-s" with a r-^nge of 3.1-13.3 ug-i; . 
Pocertlal toKiclty in food chains. 

The detailed comparison of uei lands ^t Point Anne .ind Hay Bay showed 
that As Is present in the irea of Molra drainage, )n elevated arauunts i'l 
■both' BE-Jtment and lubnie rged planfi. In the snails, h.jwever, higher Ab 
values were found in Hay Bay. 

Copper is high Ln the lower b^y. and did not differ In planti at Che 
two onshore areas. The .-lsvati"d Cu in Hay Bay «3S also found In anaiU, but 
,,,aly U Stagnicola elodes . The specificity of uptake was shLiwn by Che 
highest 1» values being found in the other sampled species, Planorbella 
Crlvolvis . 

So Far, our r-jiearch iodic-ites that of the seven metals thought likely 
to be of concern, \s and Cu -ire raosc important in the wetlands. They occur 


in sufficiently elevaced concentr-it Ions at same sices tn the Bay to be taken 
up at elei;ated levels by plants and molluacs, and therefore passed on tn 
upiwr trophic lewele. In both plants and molluscs the patterns of uptake 
ure species - specific and astal •peclftc. 

MhSlp some oetala (notably A«> have the predicted attenuation pattern 
away from the Holra, geveral unexpected distributions have been ftmnd, such 
ds Pb ^t Belleville, Cu In Hay Bay and nlnor concentrations of several 
laetals at Creek mouths. 



Crowder, *.A. and J.M. Bristow. 198f>. Aquatic nacrophyces ol the Bay of 

Quinte: 1972-82. In Project Quince: Point source phosphorus control and 
ecosy^itMrii response in the Bay of Quint*. Lake Onlarto. ed. C.K. HinnB, tl.A. 
nurUy, and K.H. Nicholls, pp. 114-127. Can. J. Flah. Aquat. Scl. Spec. 
Publ. Bfi, 

Crouder. A. A., B. McLaiighUn, R.D. Weir, and W.J. Christie. 1986. 

Shoreline fauna of the Bay of Quinte. In Project Quince; Point source 
phosphorus control and ecosysceo response in the Bay of Quince, Lake 
Ontario, ed. C.K. Minns. D.A. Hurley and K.H. Nicholls, pp. 190-200. Can. 
Spec. Publ. Fish. Aquat . Scl . *(6. 

Crowdcr, A. A. and .KM. Bci?tow. 1988. The future of waterfowl habitats In 

the Canadian lower Great Lakes wetlands. J. Gt. Lakes Res. 16(1): 115- 
I 17. 

Crgwdtr, A. A., W.T. Oushenko, .1. Greig -rnd I.S. Poland. 1988a. 

Deturmlnacloo .if mstal concauii rat Ion in «ei: Und'i (if c'lp Bay of ijuince. ^''^'' 
OnCarlo, Canada, 'roc. Int. Conf. on Envl ronnienc«l Blonsa.iy Tpchnlque^ and 
their Application, -Tuly l!-U, 1988. University of Lancaster. In press In 

Crowdt-r, A. A., W. Duslienko rtnd J. Grein. 1988h. Met«l« and th^Ir 

dUtrlbiitlnn In wetlandu of the Bay of Quinte, I.akp Ontario. Proc. tnc. 
Conf. on Trac-^ HetaU tn Lake-f. National Water Research Instltuf, 
Borlitigton. Ontario, August 15-18, I98l». In press In Science of the Total 
Eni'i ronmen! . 

Crowdiir. A.. W.T. Dushenkii ^nd I. Gr-lg. 198fic. Shoreline distribution of 
contaminants <ind -Jubo-rrt^d mjcrophytns In the Bay of Quinte, Ontario. 
Peder^Hor, of Im-irlo Naturalists. Ptoc. Onlnrlti Vet lands Conference, 
Ryerson Poljft.-chnica 1 Institute, Toropto, 21-22 October, 198S. In 

Geii, J.W. 1985. F,nvl roiim.-ntal Influenc-'s on the distribution and 

composition nf wetlands in the r.reat Lakes Basin. In Coastal Wetlands, 
^d. H.H, Prince and K. M. D'Icrl, pp. 15-3U Chelsea. MJchiaan; 
Lewlfl Publishers. 

Olooschenko, W.A. 1978. H»diflii>nt^ry Keocheraisiry of a Lake OntarU urban 
siarsh. In Procpedings j' ?nd Wurksh'tp on Great Lakes Coastal Erosion 
and SfldinKnt.iClon. .^d. N. A. RykflyUd. CCIW, Rurllngton, Ontario. 

Larrick, W.H. and R.H. Chahr^ck. 19?'). The effects of velrs on aquatic 

vegetation along r'le Li]i.'.«land co^*t. In (td) W.A. Rogers. Proc. 13th 
Annual Conf. S.E. AssocUtron Fish and Wildlife Agencies. 24-27 
Orttrib-fr 197ii, .lackson, Mlaslsslppl. pp 581-569. 

Minns. C.K.. C.l. Owei, and •i.n. T,ihnHon. l'*8fi. Vutrleni load and budgets 
tn the Bay of Quinte, 1965-81. In Pr.tject Quinte. od. C.K. Klnns, D. 
«urUy, and K. "ilcholls, pp. 59-7^. Spec. Publ. Can, .1. Fish. Aquat. 


Scl. 86. 

Mudroch, ^. and J.A. Capobl-nco. 1979. Effects ot nine affluent on uptake 
of Co. Ni, Cu. \B, Zn. Cd, Cc. and P by aquatic Mcrophytes. 
Hydrohfologla 6A(3): 223-231. 

Modroch, ^. .nd -l.A. CapoManco, 1^80. impact .r past mining ;|"t-'^^" "" 
aqaatle s*dlm«nts In the Molca river basin, OataMo. .1. Ct. Lakes Res. 
6(2): Ui-128. 

^^..dr„ch. A.. L. Sarazln and T. Lows. 1988. Su»«ary of surface and 

bj.-k«rf,and concentrations of selected eleawnta In the Great Lakes 

fiBdlmenta. J, Gt. Lakes Res. 14(2): 241-251. 

Sttlonal Wetlands Working Croup. 1988. Wetlands oi Can^a. Ecological 
Land Classification Series No. 2'.. Knvi ronnont Canada. Ottawa. 

P.ehlke. R.. L. Maynes and V. McCuUoch. 1982 ^ ^•'g^y f -"«""^- '"'^'' 
study: Che Molra river. AU^rnatUcs 10(2.3): IZ-l". 

SW p. iS86. RBvie- of po^tglncial unvlronmental changes and ™'tijral 
i^acts m the Bay of Quinte. In C.K. Minns, D.A. Hurley and K.K. 
sTcholU (ed.) Project QuUte: polnl-soorce phosphorus control and 
ec«ns"« response in Che R^y of QulnCe, Lake Ontario, pp 7-26. Can 
Spec. Pobl. Pish. Aqu^t. Scl. 86. 


-^-jT-F * - T 

1. correlation cMffieienta for vegatitlon cover and metals. 

EfitEh siitt^l SiJUUii I^"^ ^ 

Vegetation -.461 

.«SB -.725 -1S5 

c^;;/ p-.ooi p<.ooa P<0.0Q1 P-.012 P<-<'01 

Cll Hi £b in- 

vegetation -.19 .232 -«^ -af 
cover KS MS P--002 P-.03 

■ loas on ignition (420 C) 
''in sedliMnc 



Table 2. Mean and range of element concentrations in sediment Etotn near- 
ahoca sicea (n-49l in the Bay of Quince. SD- standard deviation. 





















































119. 2 

67 3 























0. 1 




Na [li Pb V In 

* ppm ppm ppm ppm 

1.31 20.9 16.0 35.9 59.7 

0.76 10.3 11.4 19.1 15.1 

Min <0.1 3.6 0.03 43 0.05 7.0 5.C 2.3 10.0 


<0.1 127.1 3.66 1449 4.72 56. 55.0 109,2 270.0 

* levels below detection limits precluded deterrr.inat ions o£ standard 
dBviationa . 

Table 3. Correlation o£ metols in sedlnwnt with physic«l and 
cheioical factors at 49 near-shoco sites in the Bay ot Qutnte. 

Slltlll Sand(%l ClayHJ LOI^ pH^' 

(neae shore] 





















" loss on ignition (420°C) 
in sedlmant 


Table i. Compeiison of mean values of roetals in sediment at Bend 
B«y, Belleville and Hay Bay marahea (on-atioce sices) . Values in 
bcackecs are standard deviations. 






Loc scion 






Ber.d Bay 


60. S 












31. S 










Hay Bay 





89. 1 






(±25. 4 J 


Table 5. Compariaon of njetala in Myriophvllmn aoicatum and Vallianeria 
ajieri{:ana in on-shor« sices tB«llevllle/Pt . Anne and Hay Bay). 

Moan Tissue Levels 


As Cr Cu 

ug/g ug/g 07/g 





Belleville (n 


M. f^pi^ariim 









V. amerlcana 









Kay Bay (n-4) 

>L. gpicattfin 









V. ameiicana 










COPFEB (ev»i 

Fig. 1. OistiibuLiOnal patterns Ot Cu, Pb, Hi an.! Zn in sediment.s f nmi 
Bend Bay and north shoce sites from the Bay of Ouinte. Numbers on the 
hoiiiontal axis order the sites by distance troir Benci Bay: actual 
distance from Bend Bay lo site 20 is ± 90 km. 



M. Slivitaky, INRS-EAU, Ste. Foy. Quebec. 





II . Hon ison , A- Benedck , and J.J. Bancsi, Zenon Envirunmental Inc. 

and S. Beszedits 

In the past decade the potential for con tarn ination o£ 
rintario dfinkimj woter sources by toxic organic pollutants has 
Ij'^cnme Jncreas i nij 1 y apparent. Many sources have chronic low level 
contamination, otiier sources could potentially be contaminated 
with hiyh levels Cor a short time period as a result of an 
ace idental spill. As a result many common i ties in Ontario are 
"ither alreody using or are proposlny to use powdered activated 
c.irbon (PAC) . ZENON Environmental Inc. has undertaken a study for 
the Ontario Ministry of the Environment Co develop process design 
i;ritecla for carbon contacting and to undertake the basic research 
necessary to ensure that the addition of PAC, if deemed desirable, 
proceeds in nn optimum manner for all concerned. 

At the time of writing approximately ten months of a 24 
month schpilule has been completed. Initial study included a 
detailed literature review of tiie application of PAC for or'janic 
removal in water treatment and a search of the wastewater or 
process water treatment literature for PAC reactor designs with 
potential applipation for drinking water treatment. Process 
design criteria for application ot PAC in drinking water 


tieatment tot the removal of toxic oryanic contflnilnints were 
developed jnd two PAC contacting systems were selected from 
potential contactiny systems. These two contactor systems ace 
beinc] developed and evaluated in bench scale systems and the best 
system will be demonstrated at pilot scale in 1989. This paper 
suminar i zes the results of the literature review and discusses the 
process desiyn criteria and the selection o£ PAC contactor systems 
for evaluation, 


The application of powdered activated carbon (PAC) is a 
routine practice in many water treatment plants around the world. 
Approx iinjtely 90% of water treatment plants worldwide that use 
activated carbon do so in the powdered form (Fajst and Aly, 1983), 
PAC is used by most of the EEC countries primarily as an emergency 
neasure in case of temporary pollution of the raw water source 
(Water Research Centre, 1977). Large amvjunts of PAC are also 
used, particularly in the Netherlands, for taste and odour 
removal. In the United Kingdom facilities for dosing t'AC are 
available at approx imately IOC water treatment wor ks (Hayes and 
Hhitford, 1982). In recent years there has been a growing trend 
toward the installation of granular activated carbon (GAC) 
adsorption systems for treatment of low level contamination. 

PAC is usually added before coagulation or immediately 
be fore the filters. The optimum point of appl i cat ion should 
allow adequate dispersion of the carbon and sufficient contact 
time to ensure maximum adsorption. When relat ively low doses are 


required (i.e. 10 my/L or less) and tiltet runs are long enough, 
PAC is added immediately ahead of Lhe filters. Experiences have 
also demonstrated that a yiven amuunt of PAC is more effective 
when deposited on the CiLtec; however, care must be exercised to 
prevent PAC Cram passing the filter in this situation. Moreover, 
to iinpcovt* the filter's ability to retain carbon particles, a 
'^mall dose of polymer is usually added. When relatively high 
doses of PAC are required, or when filter runs are already short, 
I'iSC is added ahead of the coagulat !on-f locculat ion zone in order 
tltat as much nay be removed by sedimentation (if conventional 
treatment is employed) as possible. 

Whilr? in normal practice typical PAC doses required are 
in the range of 3 to 15 mg/L, in some instances, doses as high as 
;^-1i) Hig/I. have been utilized. The PAC dose that can be effectively 
Lised for direct or in-line filtration ,Js usually limited to 
b-tween 10 and 15 mg/L. Dosage in most water treatment plants is 
established by trial and error. When largo amounts of PAC are 
required to reduce the concentration of oryanics, adding the 
'jorbon in two steps [i.e. split dose) usually results in reducing 
the total (]iiantity of carbon required. 

Although PAC may be fed as either a dry chemical or a 
slurry, it is mote preferable to store PAC and feed it as a 
slurry. Slurry tanks are usually constructed of conctete and 
equipped with a mixer to maintain carbon in suspension. 
Volumetric metering or proportioning pumps are best to use in 
feed i ng . 



The sur«oy of published literatuce did not reveal any 
alternative PAC contacting and separating methods being used or 
developed tor drinking water treatment. However, a wide variety 
oE systems which are being jgiid or developed Eor wastewater 
treatment were idenCiEied. 

PAC treatment o£ water ot wastewater in the simplest 
sense involves the intimate contact oE the carbon with the stream 
Eor a isuCEiclently long period to attain as much removal oE 
unilesirable constituents as possible EoHowecJ by ,'jepacation o£ the 
carbon from the liquid phase to prevent carryover of carbon fines 
In the discharge and/or to recover the spent carbon for 
regeneration. Thesis two fundamental steps are illustrated in 
Figure 1. As indicated in Figure 1, contacting PAC with the 
Stream tn be treated can be achieved several ways, e.g. flash mix 
ch.imber , [ 1 u id 1 zeJ bod , pi pe reactor , etc. while carbon separat ion 
can be accompl i sheil by sedimentation, Eiocculat ion-sed imen ta t ion, 
air flotation, foam Cractionat ion, filtration , centr iEugat ion, 
etc. Carbon contact with the stream and separation may also be 
caccierl out in a single unit such as the reactor-clar 1 f ier 
depicfd in Figure 2. (ienco. It is evident frym these two figures 
that many comli ina t ions o£ PAC contact and separation operations 
.3rp possible. 

Although cacboh contacting and carbon separation 
i (ocesses can be considered individually to Identify the most 
viable system for a particular application, it Is more appropriate 
to consider tliese two bnsic operations as part of the overall 


froatment sclieitie as depicted in Figure 3. 

Based on the literature review, process knowledge and 
consultation with MOK staEf, twelve reactor systems were selected 
for initi:il evaluation. These systems are shown in Table 1. 


The reactor alternatives which wtie identified as a 
r^^ult i>E the literature survey were evaluated with respect to a 
set oE criteria which were established to ensure optimization o£ 
the adsorption process and to meet the requirements of treatment 
i;lant operators and the HOE. Table 2 summarizes tiiese criteria 
lot optimum PAC adsorption. The essential requirements are 
"Tt'ective contact of the PAC with the water stream to allow 
adsorption to occur and eEfective separation o£ the PAC from the 
w-iter to remove the contaminants after the adsorption. 

When PAC is brought into contact with the water stream 
■I ;•; solved pollutants are removed by adsorption. This is an 
"quilibrium process and thus the capacity of carbon to adsorb a 
l>-irfcicular compound In the water is dependent on the concentration 
'jf that compound in the aqueous phjse. The relationship between 
capacity (q) and equilibrium concentration (C) is given by 
isotherm data and can be expressed as: 

q = KC 1/n 
where K and L/n are experimentally determined for a specific 
compound over a certain range of concentration. 

In a plugflow roactor, the initial layer of carbon 
adsorbs contaminants until it reaches equilibrium with the 


inEluent contaminant concentration. When Clus equil ibt ium 
condition is reached, the removal capacity o£ the carbon Is 
ex ha us ted. This equilibration process progresses through the 
carbon bed as each layer reaches equilibrium with the influent 
concffntrat ion . Tliis steady exhaustion of successive carbon layers 
at the inEluent concentration rGSults in optimum utilization of 
the carbon, as the carbon is in equilibrium with tlie influent 
concentration, and very low concenfctat ions of oqei taminants in the 
produced water, as the cfEluent Is in equilibrium with virtu.illy 
unused carbon . 

The conventional PAC application In drinking water Is a 
completely mixed reactor. In this system the only adsorption 
equilibrium is between the carbon and tlte final effluent 
concentration. Thus the carbon is not used efficiently and the 
product water has a relatively high concentration of contaminants. 
In a reactor where the PAC contact Is not plugflow if two or more 
reactors can be used in series and the PAC moved counter currently 
through the series PAC adsorption capacity and effluent quality 
can be optimized. 

Thus an essential criteria for selection is that contact 
be stratified (plugflow or mul ti staged) . 

In any reactor PAC contact Llrao mu.'iL be optimized to 
maximize adsorption. Kinetic studies have In the past focused on 
granular activated carbon. For granular carbon adsorption optimum 
contact time is generally greater than five minutes however as the 
particle size decreases optimum contact time decreases and thus a 
contact time of one or two minutes may be sufficient for powdered 


activated carbon. For selection cr iter ia , one minute was used, 
however, experimental testing ia required to further deEine this. 

The third essential criteria is that the PAC he 
adequately retained by the reactor to prevent deposition of PAC 
downstream in tlie water treatment process or distribution system. 

Another essenti.Til requirement for the PAC system is that 
it must be able to adsorb occasional high levels of contaminants 
in rosjjonse to a chemical spill in the water source. This 
capacity may be automat ic or in response to adjustment oE 
operatinij conditions by operator. 

Other essential crifitia are tliat the system be 
adaptable to existing facilities, the process mjst operate 
reliably and the system must be mechanically reliable. 

The "desirable" criteria were included in this analysis 
as conditions which would have benefits for the MOiv or treatment 
plant operators but were not necescacy to an improved PAC 
contactor system. The first "desi c ablo" criteria was that the 
system he portable so that it could be moved to a facility 
encountering temporary contaminant problems. The second desirable 
criteria is that the PAC be recovered in a form suitable for 

The third class of criteria were the cost ciltecia which 
were included to ensure that any system selected for further 
invest igat ion have reasonable capital, labour, and opera t ion and 
maintenance costs associated with it. 



The twelve PAC systema were evaluate'! with resppct to 
the established criteria using inEormation obtainer:] from the 
litoratuie. Information obtained in bench scalt tefitinq and best 
engineering judgement. Four of the systems met the essential 
criteria. Those were 1) Solids Contact Clarifier, 2) 
volat ilizat lon/Gaa Phase Adsorption, 3) Down Flow Filter and 4) 
Multistage CrossElow Filtration. With respect to the desirable 
criteria it was uncertain if any of these systems could be made 
readily portable. All systems except the solids contact clarifiec 
would produce exhausted PAC In a form suitable for regeneration. 
With respect to costs, the solids contact clariEier is expected to 
be the least expensive alternative, while the other systems would 
depend on innovation a nil optimization to bring the costs to an 
acceptable level. 

For the purposes oE this study, the downflow filter and 
the multistage ccossflow filter were chosen for further study. 
Tlie solids contact clarlEier h-is been well studied for wastewater 
tre.itment and is bijing optimized in on-going studies by V. 
Snoeyink {Snoeyinh l^fla). The vol j t i 1 i z*i t ion/yas phas-? adsorption 
system is limited Ln application to compounds which are 
efficiently air strippable .ind thus w.33 f^llminated from further 
study on this basis. 

Bench scale testing of PAC downflow filtration and 
multistage crossflow filtration Is currently on-going. A 
technical and economic evaluation of these two systems will be 
made based on the results of these test. 



1. Faust, S.U. aniJ O.H. Aly (19liJl Chf?iii;stry af WatT Treatment. 

Buttotwoc ths, Boston, HA. 
a. Hayes, C.S. and C.J. WhitEocd (1932) The Use oE Activated 

Carbon in Water Treatment, KEEluqnt a nij | ^a ter TrgatmpiU T. , 

Vol. 22, No. 1, pp. 9-16. 

3. Snnr-yink, V. (1988) Personal Communications. 

4. Water H^searcli Coritce (1977) Wator Purification in the r:i;C, 
Pergamoii Press, Oxford, Great Br i tain . 



■neu 1 



ConUcting Syst^ Sepaiating Systai 

Separating System 

Sand Filtration 


Flota t ion 

Solids Omtact 
Clarif ier 

Volatilization Gas 
Phase A^socption 

Down Flow Filter 
Up Flow Filter 

Fluid i zed Bed 

Dead End Surface 

Surface Filter 

Centr 1 tuqe 

CrossElow Surface 

Rapid Mix 
Rapid Mix 
Rapid Mix 

Saiul Filtor 

coagulation Sedimentation 


Solids contact 

ftd sorption 

treatment of 
vapour from air 
atr ipper 

Packed PflC colurr 

Column of PAC 
attached to 


Up Flow thcouyh 
fluidized PAC b<?i 

Rapid Mix 
Rapid Mix 
Rapid Mix 

Plate and fratne or turous 
tube filter 



Counter current 
flow of PAC 
through a series 

of Ultrafiltration 



A) OpttiBum Contact Time 

R} Contact StratiCication 

'■) Holiable Separation 

D) Spill Response 

K) Adaptable to Existing Facilities 

y) Process Reliability 

G) Mechanical Reliability 

|[) Poctab i 1 i ty 
I ) PAC Recovery 


Jt Capital Cost 

K) Labour 

l..| Operation an^i Mii in ti^nancft 




fresh PAC 

raw feed 




treated water 



flash mix tank 
oipe reactor 

spent PAC 

fluJdized bed 


foam fractionation 
air floatation 



raw teed 

fresh PAC 

Reactor - 

treated water 

spent PAC 




(resh PAC 

raw feed 

prior to 
PRC contact 





spent PAC 




' ' 








R. D. Kreuczvlser 

R. B. Feagan 

Depattment o£ Geography 

University of Guelph 
Guolph, Ontario, NIC lUl 

For presentation Co the Environmental Research^ Tttchnology Transfor 
Conference , Toronto. Ontario. Hoventbor 28, 1938. 



The mflnagemBnt of municipal uaEer supply has buen dominated by a supply 
aanagenent philosophy, which has stressed expanding supply to mocC demand 
CGovernmont of Ontario. 1984). However, economic and environmental 

realities Increasingly call into question the appropriateness of this 
philosophy and suggest the viability of dematid management (conservation) as 
an alternnCtve or supplement to traditional supply management. Recognition 
of this comes from senior governments In Canada (Ministry of Natural 
Resources et al.. 1985; Canada, 1987), though present use and promotion of 
water conservation strategies appear limited. 

This paper describes the extent of use of demand raanageraent strateglea 
among southern Ontario municipalities, discusses several factors thought to 
influence municipal adoption of consfirvation strategies, and suggests how 
munleipalUies can enhance their conservation efforts. 


A questionnaire survey was sent to the water system managers (Publ Ic 
Utilities Commission general manager, city engineer or ctjuivalcnt) of 315 
southern Ontario municipalities with public water supply systems. The survey 
was designed to obtain Information on use of a broad range of conservation 
strategies, as well ss characteristics of the water system and water system 
manager thought to influence adoption of demand management. 

Useable responses were obtained from 219 municipalities, or 70* of 
those surveyed. This represents a population of about 6.300,000 of the 
7,000.000 people served by municipal water systems In southern Ontario, 


Characteristics of Water Syscanis 

An Impression of the municipal water syatems represented by respondencs 
can be gained by considering the following facts. Almost 53 percent of water 
systems were council-run with 39 percent operated by s Public Utillcles 
Coraitssion. Fifty-five percent drew from a surface water source while Ui 
percent pumped from a groundwater source. 

The eaphaals on supply nanagenent is evidenced, in part, by Che fact 
thac in the last ten years, 35 percent of systems expanded pumping capacity 
and 30 percent expanded storage capacity. Fifty -one percent of those on 
groundwater Hddod uella and 10 percent of all responding systems added 
treatment capacity. Problems are evident . however. In that 36 percent of 
systems encountered summer shortages. 36 perecenc experienced water quality 
problems and 32 percent had major distribution leaks during the past ten 

Utilization of Demand Management Strategies 

Table 1 reports the extent of municipal utlllEatlon of a wide range of 
demand management strategies. 

In terma of economic strategies, only three municipalities made any use 
of a weCsred inclining rate, which is considered a very powerful conserving 
strategy. Thirty-six percent made some use of a metered flat rate, while AC 
percent used a metered decliniing rate. A set price rate structure was in 
place In almost 60 percent of aystena, While attaching a price to water use, 
chls strategy has no real conserving Influencing. Forty-eight percent of 


responding nunlclpalities Iraposed a seuer surcharge and 54 percent Increased 
races in 1987. 

As reflected in Table 1. use of water metering is extensive; 79 percent 
of nunlcipallcles had metering In place for at least sooie use sectors. 
Almost half had some neCering of residential users, and extent of metering 
was higher In other sectors. Only a minority of responding municipalities 
spent more on meter installation and repair and distribution system leak 
detection and repair in 1986 than the mean of their expenditures over the 
previous five years. 

Municipal use of regulatory strategies was generally lov. Thirty-five 
responding municipalities have imposed summer use restrictions and 22 
percent have used voluntary restrictions on water use . Only 17 
municipalities had water-conserving plumbing ordinances lin place. 

Educational strategies were also little used, with only 22 percent of 
municipalities distributing educational material with water bills. Fewer 
(six percent) made any attempt to educate major commercial or Industrial 
water users. Only 6 municipalities distributed retro- fit water -conserving 
devices In an effort to encourage water conservation among residential 
users . 

Several water conservation sCrataglas described in the muntclpal water 
conservation literature were not In use among any of the responding 
municipalities. These were dally peak-hour rate structure, marginal pricing 
for new water users, and Llnlcing distribution syscen pressure to a mininuiD 
acceptable level . 


Influences on Utllliatlon 

Several chacacterlBtlcs of the water system and water system manager 
vera hypocheslzed as influencing municipal adoption of deitand management 
stracflglea, Chi -square analysts was used to tesc for asaoclaclons , It was 
found chaC greater use of water consarvation strategies was associated with 
larger populations seved, older distribution system age , greater extent of 
water problems experienced. Involvement of regional municipalities as water 
suppLlcXB. and systems operated by a Public UcLlltles Commission. It was 
thought that use of water conservation strategies would be associated with 
nunlclpaliriea drawing from a groundwater source, rather than a surface 
water. However, generally the opposite was so, which nay reflect the 
increasing costs of tretlng surface water. Ko association was found between 
adoption of water consarvaclon atracagles and experience and training of 
water system managers, 

Many of Che factors outlined above axe interrelated, and further 
analysis was conducted Co isolate the most influential Independent factors, 
Ic appears that population served and extent of problaas are moat strongly 
associated with municipal adoption of demand management acrategies. 

Enhancing Demand Hsnagananc 

Hunlcpal water rnanagemenC literature suggests that demand management 
can reduce supply and treatment costs, reduce waste water treatment costs, 
off • set tamporairy supply shortages , reduce water appropriation conflicts, 
minimize environmental costs of system expansion, and defer system expansion 
tEllls, 1978; Haier et al,. 1981; Sawyer, 198J: Grlma, 1985), Despite these 


reporcad benefits, municipal adoption of ch« demand nanagenent concept: anong 
southern Ontario cnuntcipaltties has not been extensive. 

There are . however , several actions municipalities can take to enhance 
water conservation. Hunlclpalltles, for example, can move toward full 
metering of all water users. While It can he debated whether meterlnlg by 
icselE has a major conserving influence, users becone avare of the volume 
used and they pay In proportion to use. Loudon (1984) found a 13-20 percent 
decline inw ater use In Durham Region after Introduction of metering. 
Metering Is also a basis for applying rate structures that can influence use 

Mxinlclpaliciea can also nova «way from a declining rate structuce to 
more conserving structures . While the flat metered rate Is less effective 
than the Inclining metered rate. It is more equitable and tmplemenCable. 
General rate Increases, to better reflect the full costs of water supply and 
the value of the resource, can also influence water use , Several studies , 
for example Ellis (197S). suggested an average absolute price elasticity for 
municipal water of about .5, which means that a price increase of ten 
percent would result In a five percent reduction In use. 

Hunlclpalltles can increase effort to detect and repair distribution 
system Leaks, MacLaten (1985) estimated 28 percent unaccounted for use among 
Ontario inunicipalitles and 35 breaks per 100 kia per year. Under these 
circumstances, leak detection and repair could be cost-effective. 

Barclay (1984) suggested that water -conserving devices on plumbing 
fixtures can reduce water use by over 30 percent. Plumbing and building 
ordinances, though presently little used In Ontario, offer the potential to 
reduce consumption substantially. Limiting distribution system pressure to 


nlnlauni levels acceptable for f ire-fighclng purposes can slso reduce use and 
leakage, though this reduction may not be great, 

Education Is usually considered to have minimal Impact on water use 
behaviour, while econoalc strategies appear to offer greatest potential. 
However, education can have an Important role In enhancing the local 
BccepCablllty and adopclon of more effective strategies. Edcucatfon can also 
play a role in developing political support for changes in senior governnent 
grand structures and other arrangements Influencing local decision-making, 
to ensure that appropriate Incentives for demand management are established. 

The financial support of che Ontario Kinlstry of the Envltonnenl: and 
the assistance of Philip Joseph of the Ministry are gratefully acknowledged. 


Barclay. D.S. 1984. Retrofitting apartment buildings co reduce casta 
and water demand, Canadian Water Resources Journal 9(3): U^-itl . 

Canada. 1987, F^ij^rq^ tf ^t^^ r PpUtJY - Eiivirotimerc Canada, 

Ellis. R.K. 1978. New considerations for municipal water syscen 

planning. Water Resources Bulletin 1-^(3): 542-553. 

Gf>vernment of Ontario. 19BA . Futm f^ B jq Water . Toronto. 

Grlma, A. P. 1985. Urban water conservation, p ^Q^ pu rnB l , U(3): 357-263. 

Loudon. R.M. X^^h . Region of Durhajn experiences in pricing and water 

conservation. Canadian Uace;- Resources Journal 19<i) : 19-28. 

MacUren. J.W, 1985. HOTli;tp^1 Wqtgr Woyks and Wastewater Systems. 
Federal Inquiry on Mater Policy Research Paper 3, Ottawa, 

Maier. V.J., DeZeller, J. and Miller, W.L. 1981. Benefits from water 
conservation depend on comprehensive planning. Water Resources 
Bulletin 17(4): 672-676. 


Ministry of Natural Resources. Ministry of the Environment and Ministry 

of Agriculture and Food. 1985. Towards a Water ConservacLon 

StrHtzegv for Ontario. Toronto. 

Sawyer, S.W. 19B3. Water conservation; conflicting attitudes of 
planners and utility nanagers , T^e Environmental Professional 5: 


Tabla 1 Use of Uater Conservation Stracegies Among Southern Ontario 

Percent of 
SCrftCegy Municipalities 

Race scructurn 

• set price 58.9 

■ atetered declining 40.2 

- metered flat 36.1 

- metered inclining 1 ■* 

Overall rate Itw:rea«e (1987) '•■2-'^ 

Sever surcharge 6'*. 8 



■ any sector 

- residential '■93 

- conoierclal 68.9 

■ Industrial 60,3 

- Institutional 52-9 

19S6 Metering budget greater than mean 11.9 
of previous 5 years 

1986 Distribution system leak budget greater 13.7 
Chan mean of previous 3 years 

Sunmer u;»e restrictions 34.7 

Voluntary reatrlccions 21.6 

Pluablng and building ordinances 7-8 

Uater-conservliig devices 2.' 

educational inictatives 

• pamphlets 21. 5 

- nedia Inforaation 12. B 

- troat»enC plant tours 17.4 

■ commercial/industrial prograres 6.4 



A Preliminary Study to Determine the Feasibility 

of Medium Pressure Mercury Lamps for Disinfecting 

Low Quality Wastewaters. 

G.E. Whitbvi. J. Haarschallterweerd, , ana 
G.A. Palmateer, 

1. Trojan Technologies inc. 

845 Consortium Court 
London, Ontario 
N6E 2S8 

2. Ministry of the Environment 
985 Adelaide Street South 
London, Ontario 

N6E 1V3 



During dry weather moat wastewaters receive some form of 
chemical or biological treatment to remove organic and inorganic 
constituents and then these effluents are disinfected to protect 
users of the receiving waters. It is only during periods of 
rainfall that significant quantities of eflluents such as 
stormwater runoff and combined sewer overflow are allowed to flow 
Into watercourses with little or no treatment. Combined sewer 
overflow is the result of joint wastewater and surface runoff 
collection systems . These combined sewer systems are prevalent In 
older areas of most municipalities. These microbiologically 
contaminated waters have contaminated raw water supplies and 
swimming areas throughout North America. The closure of swimming 
areas is a great inconvenience for everyone and results in a loss 
of revenue for those involved in tourism. 

Programs to alleviate the situation include the separation of 
combined sewers which Is very expensive, methods of reducing the 
volume and frequency of overflows, and methods of improving the 
quality of the storm runoff. It is this latter aspect where the 
use of medium pressure mercury ultraviolet lamps was investigated 
lor the purpose of disinfecting low quality wastewaters. 

Previous studies (Scheible, 1985 and Zultovs, et al, 1986) 
have shown that combined sewer overflow (CSO) can be disinfected 
with low pressure mercury lamps but the capital cost of the system 
Is very high compared to chlorine because of the large number of 
lamps. The high cost is a result of the high flow rates and long 
retention times which are required to obtain a three logarithm 
reduction in the number of fecal conforms. 


A medium pressure mercury lamp has a much higher intensity 
per unit of arc length compared to a low pressure mercury lamp. A 
low pressure mercury lamp which is normally used (or the 
disinfection of liquids has a UV output oE approximately 0.2 watts 
per centimeter o£ arc length at a wavelength of 254 nm. The 
output of UV light is almost monochromatic and within six 
nanometres of the optimum wavelength for germicidal action. 
Medium pressure mercury lamps have an average UV output of 9 watts 
per centimeter of arc length at wavelengths below 360 run. 

If all of these wavelengths below 380 nm were equally 
effective at killing fecal collfomts. a medium pressure mercury 
lamp would have 45 times as much germicidal power per centinieter 
of arc length, A significant decrease in the number of UV lamps 
would result in a much lower capital cost which could make UV 
Irradiation of low quality wastewaters an economically viable 
process . 

The study had three phases. The first phase determined the 
dose of UV light which was required from a low and medium pressure 
mercury lamp to disinfect a series of different quality 
wastewaters. The second phase looked at the total UV output of 
low and medium pressure mercury lamp so that a pilot system could 
be built. The third phase involved the testing of a medium 
pressure lamp system with three different waterlayers to determine 
the economic feasibility of the process. 



1. Purpose 

This phase oE the study compared the monochromatic light (254 
nm wavelength} oC the low pressure mercury lamp to Che broader 
spectrum ot the medium pressure mercury lamp. The bloassay method 
of Quails and Johnson (1983) was used to ma)c.e this comparison. The 
bloassay uses a collimated beam of light Co irradiate a volume of 
stirred wastewater. 

Two collimated beams were adjusted so that a UV sensor which 
was only sensitive to light around a wavelength of 254 nm showed 
that the beams were equal in UV output. The additional 
wavelengths in the spectrum of the medium pressure lamp should 
show up in the survival curves of the fecal conforms in the 
various wastewaters . These survival curves can then be used to 
compare the two types of mercury lamps. The dose of UV light was 
adjusted so that a three logarithm kill or 200 fecal coliforms per 
100 mL was reached in every wastewater. 

2. Materials and Methods 

Samples of raw wastewater after the comminuting devices, raw 
wastewater after primary settling and secondary treated wastewater 
were obtained from the Greenway wastewater Treatment Plant in 
London. Ontario, Canada. Samples of raw water after the 
comminuting devices were also obtained from the wastewater 
treatment plant in Ingersoll, Ontario, Canada. 


Various strengths of cso were prepared by mixing secondarily 
treaced wastewater so it contained 13.5, 25 and 50 percent raw 

The bioassay method of Quails and Johnson (1983) was modified 
so that the medium and low pressure mercury lamps could be 
compared. Two collimated beams [Figure 1) were set up side by 
side so that the same day's samples was irradiated with the medium 
and low pressure mercury lamps. 

Botn light sources were set at 200 microwatts/cm' at the 
liquid surface with an International Light 1500 Radiometer with an 
SEE 240 sensor (International Light Inc, Dexter Industrial Green, 
Newburyport, Massachusetts. USA) . 

A single (SOmL) of wastewater was measured into the 
irradiation chamber shown in Figure 1 and continuously stirred 
during tne exposure to UV light. A series of exposure times was 
used for each wastewater. The wastewater was 2 centimeters deep. 

Unirradiated samples were stirred to determine whether the 
suspended solids were being brolcen up by the magnetic stirring 

Eacn sample of wastewater was analyzed for UV transmittance 
at a wavelength of 254 nm, Each wastewater was filtered through a 
0.«I5 micron Gelman GN type filter and analyzed for UV 
transmittance at a wavelength of 254 nm. 





Figure 1; Schematic diagram of Che collif^iaced beam apparatus for 
irradiacing the various wasCewaCers. 

The total suspended solids of eacn wastewater was analyzed 
according to Method 209C In the 16th Edition of Standard Methods 
For The Examination of water and Wastewater [American Public 
Health Association, 1985) . 

The fecal coll forms were measured by the membrane filtration 
and rota-plate method. The membrane filtration method and the 
media for the rota-plate method were from the (Ontario Ministry of 
the Environment, 1984) Handbook of Analytical Methods for 
Environmental Sanplea. 

Each wastewater was tested at least five times or until 
consistent results were obtained. 

3. Results and Discussion 

The results in Table 1 show that stirring the raw. primary 
and secondarily treated wastewaters in the irradiation chambers 
had no effect on the count of the fecal conforms. Therefore, the 
samples can be stirred during the irradiation without breaking up 
the suspended solids. This is important because the size and 
level of suspended solids affects the degree of disinfection which 
can be attained (Quails et al. , 19851. 

Figure 2 to 8 and Table 2 summarize the paired testing of the 
low and medium pressure mercury lamps on the raw. primary, 
secondary and mixtures of wastewaters. 

All of the kill curves are typical of that found for fecal 
coliforms in wastewater (Quails et al. , 19B5) in that a final 
plateau is reached where increases in the dose of UV light has 
very little effect on the level of fecal coliforms. This is due 
to the suspended solids which protect the fecal coliforms from the 
UV irradiation. 


Table 1: The efface o£ sctrring the wascewaceT in the irradiation chambers on Che 
level of fecal collforms. 

Ceofficcric Mean Fecal Coliforms per lOQ wl 
Uascewacer Time in Minutes 


Raw 1 . 2 X 10^ 1 . 3 X lO" 

Primary l.U x lO^ 1.5 x 10* 

Secondary l.i x lO* l.i x 10^ 



30 40 50 60 70 

DOSE (mW, sec/mL) 


figure 2; Paired Cescing of the low {il3 and mediun (•] pressure mercury 

lamps on Che raw eifluenc frorn Che Creenway Wascewacer Treatment: 
Plane using the colllmaced beaai. 



30 rfO 50 60 70 

DOSE (mW.sec/mL) 




Figure 3: Paired cesclng of che low ( a ) and media-n (• ) pressure mercury 
lamps on che [irtmary effluenc from che Creenway wastewater 
CreaciMnt plane using che colUmated beam. 



10 15 20 25 

DOSE (mW.sec/mL) 

Figure U: Paired testing o£ the low ( • ) and medium {& ) pressure 

irtercury lainps with Che colligated beam on the secondarily 
Created effluent from the Creenway wascewater treatmenC 




30 40 50 60 70 

DOSE (mW.sec/mL) 


90 100 

Figure 5: Paired testing of the low (a) and nedluni {• } pressure mercury lai^ with the 
collimaced beam on the mixture of 12.51 raw effluent and 87.51 secondary 

effluent from the Creenway wastewater treatment plant. 




30 40 50 60 70 

DOSE (mW.sec/mL) 


90 100 

Figure 6: Paired testing of the low { A ) and nediuti {• ) pressure mercury lamps "iCh 

Che coUlMCed beao on Che mixture o£ 25Z raw and 75Z secondary eCfluenc tro- 
che Creenuay wastewater treaCment plant. 



20 30 40 50 60 70 80 90 

DOSE (mW,sec/mL) 

Flgjre 7: Paired testing of the low [a 1 and medliui { • ) pressure mercury la«ps irflth 
the colllnaced bean on the nLxCure oE 50t raw and 501 secondary effluent 
from the Creenuay wastewater treatment olant. 

■10 20 

30 40 50 60 70 

DOSE (mW.sec/mL) 




Figure 8; Paired Msclng of Che lo« (a ) ind tiediudi [ • ) preisure 

mercury lamps with the coUlmated bean on che r«H effluent 
from Che Ingersoll uastewater Creatnenc plant. 

Table 2: The ftlcered arxi unfllcered UV cransmlccartce and cocal suspended solids of 
Che various wascewacers and mixtures of wasCewacers. 


% Transmission 

1 cm 

Tocal Solids 


























12.5% Raw 

87.5% Secondary 







25% Raw: 

75% Secorriary 






507. Raw: 

50% Secondary 






Raw from 







In these experiments. Che maximum dose of UV light £rom 
the low and medium pressure mercury lamps was able to reduce the 
count of fecal conforms In the primary, secondary mixtures of 
wastewaters to below 200 fecal coliforms per 100 mL. The level of 
fecal coliforms in the raw effluents from the two wastewater 
treatment plants could not be reduced to 200 fecal coliforms per 
100 mL. The high suspended solids (411 tng/L) and low uv 
transmission (1,1%) of the raw wastewater from IngerBoll accounts 
for this difference. The suspended solids accounts for this 
difference with the raw wastewater from Greenway because the UV 
transmission oE the filtered and unfiltered samples were almost 
identical for the raw and primary effluents. 

As the wastewater proceeds through the Greenway wastewater 
treatment plant, the unfiltered UV transmission increases by 2.6 
times whereas the filtered effluent only increases by 1 . 2 times. 
The level of suspended solids decreases nine fold as the 
wastewater goes from the raw water to the end of the secondary 
treatment. Therefore the increase in UV transmission is primarily 
due to the decrease in suspended solids. 

Curves of the survival ratio versus the dose are shown in 
Figures 9 to 15 for the various wastewaters. From these curves 
and Figures 2 to a, the dose of UV light can be obtained which 
produces a three log decrease In the count of the fecal coliforms 
or reduces the level of fecal coliforms to 200 per 100 mL. These 
doses are summarized in Table 3. This table shows the quantity of 
UV light from a medium or low pressure mercury lamp which is 
required to bring the level of fecal coliforms in a volume of one 
mlllilitre down by three logs or to 200 per 100 millilitres. The 
dose of UV light varies quite dramatically between the different 
wastewaters. This is a result of the differences in the level and 
form of the suspended solids and the UV transmission. The flow 
rate can be calculated using these values for the dose and knowing 
the total UV output of the lamp. The total uv output of the two 
types of lamps was determined in Phase 2. 


10 20 30 40 50 60 70 tSO yu 100 

DOSE (mW.sec/mL) 

Figure 9; Log survival curve oE Che fecal coU£or«is in Che raw effluenC from the Creenway 
vasCewflCer treaCnenc planC aEcer exposure Co the colllMCed beam fron the low 
(A) and medii.m (B) pressure mercury lamps. 




90 100 

30 40 50 6b 70 

DOSE (mW.sec/mL) 

Figure 10: Uig survival curve of the fecal conforms In Che primary effluent fro« 

the Creenway wascewater Creacnenc plant after exposure to the colUnated beai 
frop Che low (A) and mcUub (B) pressure nercury lamps. 

10 15 20 25 
DOSE (mW. sec/mL) 

30 35 

^^"^ "' S«^^r'^ '""^ °^ "^ ^«^> conforms in the secondary effluent 

80 90 

Figure 12: 

TO 20 30 40 . 50 60 70 

DOSE CmW,sec/mL} 


O 102030 40 506070HU9U100 

Figure 13: Log survival curve oE Che fecal collforns In Che mixture of 25Z raw and 751 Becondary effluent 
frow the Greenway wastewater Creatnent plant after exposure to Che colllnaCed beaa fro« the 

low (Al and tiedlum (B) pressure mercury Latnps. 

Figure W; 

30 ^0 50 60 

DOSE (mW. sec/mL 

low (A) and medlun (B) pressure iiercury laops. 





20 40 60 80 

DOSE (mW. sec/mL) 


Tigure 15; 

L>g survival curve o£ Che fecal coUEorms in the raw effluent 
from the ItigersoU wastewater treaCmenC plant after exposure to 
Che low lA) and medium IE) pressure mercury lamps. 

Table 3: The dose of UV iLghC Cron Che nedlun or low pressure nercury laip uhlch 
produces a three log decrease in the count of fecal colLforms or rediKres 
the count of the fecal collforms to 200 per 100 nl. 


Fecal Colt form 

Dose (BiM.sec.mL ^) 

Medluai low 

Pressure Pressure 

Ldnp Unp 


3 Log Decrease 
200 per 100 nL 

13 il 

Not Reached 


3 Log Decrease 

200 per lOO mL 



3 Log Decrease 

200 per 100 mL 

Not Reached 
6 25 

12.51 Raw:87.51 Secorxiary 

3 Log Decrease 

200 per lOOoL 


251 Raw; 75t Secondary 

3 Log Decrease 

200 per 100 nL 



501 Raw: 501 Secorelary 

3 Uig Decrease 
200 per 100 nL 




Raw IngersoU 

3 Log Decrease 

200 per lOOmL 

^9 102 

Nat Reached 

The ratios of the dose of UV light from the low and medium 
pressure lamps for the various wastewaters are shown in Table 4. 
For Che Greenway wastewaters and mixtures of wastewaters the ratio 
is 3.6 (Standard Deviation = 0,5) and it ia consistent between the 
various types of wastewaters whereas the ratio for Che raw 
effluent from Ingersoll is 2.1 Because the medium pressure 
mercury lamp has a much broader spectrum of germicidal light, this 
ratio may be very specific for each effluent depending upon its 
absorption speccrum. The raw effluent from Ingersoll has a very 
high iron contenc due to the presence of a wire manufacturer. 
Iron readily absorbs UV light from ctie low and medium pressure 
mercury lamps. 

4. Conclusions 

1. Stirring Che irradiation chambers had no effect on the level 
of the fecal conforms in the raw, primary or secondary 
effluent from Che Greenway wastewater Creatmenc plants. 
Suspended solids were not being broken up so the results of 
the Irradiation should not be influenced by the release of 
fecal coliformE from particles. 

2. Suspended solids in the Greenway effluents have the greatest 
effect on changes in the UV transmission as the wastewater 
proceeds through the plane. 

3. Each type of wastewater requires a different dose of UV light 
Co reach the required level of disinfection. This is a 
result of the UV transmission and suspended solids and the 
relationship of the fecal conforms with these particles. To 
properly size a UV system for CSO, a series of survival 
curves should be prepared using the meChod described in this 
report . 

4- When each watt of UV light from a medium pressure mercury 

lamp was measured with an IL 1500 RadlomeCer with a SEE240 
sensor, it was equivalent to 3.6 watts of UV light at a 
wavelength of 254 nm from a low pressure mercury lamp when 
Che wasCewaCer was from che Greenway wascewater Treatment 


Table 4: The ratio of the doses of UV llghc from the low and nedium pressure 

■ercury laops uhich were retired to reach a three log decrease or 200 

fecil collforiBs per 100 aL. 

Ratio of 



Pressure Lanp 

MedLun Pressure Lanp 


Three Log 
Decrease In 

Fecal CoUtorms 

200 Fecal 
Collforns per 











12.51 lUw: 87 

.51 Secondary 



251 Raw: 751 




501 Raw; 501 




fUw IngersoLl 




1. Purpose 

The objective of this phase of the project was to determine 
and compare the total germicidal output of the low and medium 
pressure lamps. This would allow an economic comparison of the 
two lamps and it will also provide the information which is 
required for the design of Che medium pressure reactor vessel in 
Phase 3. 

2. Materials and Methods 

The point source summation method of Quails and Johnson 
(1983) was used to determine the UV output of the low and medium 
pressure mercury lamps. The intensity was measured at 150 cm from 
the centroid of the lamps. 

The low pressure mercury lamps were from Voltarc Tubes Inc. , 
type G36T6L. Three of these lamps were burned for 100 hours 
before measurements were taken. 

One of the 2000 Watt medium pressure mercury lamps were from 
Voltarc Tubes Inc. Two of the 2000 Watt medium pressure mercury 
lamps were from w,c. Heraeus. 


3, Results and Discussion 

The UV light output of the low and medium pressure mercury 
lamps are summarized in Table 5. The average output of UV light 
by the 2000 W medium pressure mercury lamps as measured by the 
point source sunmiation method with the SEE240 sensor is 52.2 watts 
(Standard Deviation = 1.9). This can be converted to the 
germicidal power of the low pressure mercury lamp by multiplying 
by 3.6. The results from Phase 1 showed that each watt of UV 
light from the medium pressure mercury lamp was equivalent to 3.6 
watts of UV light from a low pressure mercury lamp when wastewater 
from Greenway was being disinfected. The 2000 watt medium 
pressure mercury lamp has Che equivalent of 183 watts of UV light 
from a low pressure mercury lamp. 

The average output of UV light from the low pressure mercury 
lamps (G36T6L) after 100 hours of burning was 13.2 watts (Standard 

Deviation = 0.5) as measured by the point source summation 

One - 2000 watt medium pressure mercury lamp is equivalent to 
14.2 low pressure mercury lamps {G36T6L) when effluent from 
Greenway was being disinfected. 

The total output of Che two types of lamps as measured by the 
point source summation method can be used to calculate the flow 
rates for the various wastewaters. These flow rates are obtained 
by dividing the total UV output by the dose of UV light required 
per millilitre of effluent and then converting the answer to 
litres per minute. 


Table 5: UV ll^E ouCpuC of Che medium and low pressure mercury lamps iS neasured 
by Che polnc source sunnacLon method. 

Un^ Type Races 

Medium Pressure 

hferaeus 1 33.6 

Heraeus 2 53.9 

Vol care SO.l 

ItM Pressure 

C36T6L #1 12. t 

f 2 !}.» 

# 3 13.5 

k. . 

The flow races are shown in Table 6 for the various 
wastewaters. From this Table it can be seen that the flow rates 
are extremely variable between the various wastewaters. This is a 
result of the variations in suspended solids. UV transmission and 
the inlTLial numbers of fecal conforms. This data on flow rates 
for a 2000 watt medium pressure mercury lamp will be used to build 
a UV unit with three different water layers. This UV unit is 
described in Phase 3. This UV unit will be tested with raw 
effluent at flow rates of 482, 241 and 120 litres per minute and 
with primary effluent at flow rates of 1044. 522 and 261 litres 
per minute. 

4. Conclusions 

1. Using the point source summation method, the 2000 watt medium 
pressure mercury lamp produced 52.2 watts of UV light at 

254 nm and the low pressure mercury G36T6L lamp put out 13.2 
watts of UV light at the same wavelength. 

2. The flow rates for the various wastewaters are quite variable 
due to the differences in suspended solids, UV transmission 
and numbers of fecal coliforms. Each wastewater should be 
characterized before being treated with UV light. This 
characterization should include: suspended solids, UV 
transmission, level of fecal coliforms and a survival curve 
of the fecal coliform.s. 


Table 6: Vr^e flow races of che low and mediun pressure lamps for Che various 


Fecal Coliform 

Flow Race (L/mln) 







Not Reached 










12.57. Raw; 87.57. SecoT>dary 

251 Raw: 75^ Secondary 

501 Raw: 501 Secondary 

Raw Ingersoll 

3 Lc^ Decrease 

200 per 100 mL 

3 Log Decrease 
2OT per lOO n>L 

3 Log Decrease 
200 per 100 mL 

3 Log Decrease 
20O per 100 niL 

3 Log Decrease 
200 per iOO mL 

3 Log Decrease 

200 per 100 «iL 

3 Log Decrease 
200 per 100 mL 

















Noc Reached 


1 . Introduction 

Liquids with a low UV transmission and/or high suspended 
solids must be thoroughly mixed as they pass through a UV unit so 
that each microorganism is subjected to a maximum dose of UV 
light. Table 2 shows that raw and primary effluent have a low UV 
transmission and high suspended solids compared to secondarily 
treated wastewater. A UV unit containing a 2000 watt medium 
pressure lamp was built with three different water layers to 
determine the effect of water depth and flow rate on the 
disinfection of raw and primary wastewater. 

2, Design Specifications and Rationale for the Continuous Flow 

Of the wastewaters examined during Phase 1 of this study, two 
were chosen for the continuous flow study and these were primary 
and raw effluent from Greenway Wastewater Treatment Plant in 
London, Ontario, Canada. These effluents would represent low 
quality wastewaters with and without sedimentation. 

The proposal for the project called for mechanical mixing of 
the wastewater as it passed by the UV lamp but because of the high 
flow rates this was substituted with three different water layers 
around the UV lamp. The UV unit was built with a long inlet suid 
smooth surfaces and curves to minimize the dead corners as the 
wastewater passed from one water layer to the next. The three 
water layers were built as one unit to minimize the effect of 
having three different UV units. The water layers could be 
studied at the same flow rate in rapid succession to reduce the 
effect of changes in the wastewater quality. 



The UV unit wag built around the 2000 watt medium pressure 
mercury iamp which was used in Phase 1 of this study. This 
eliminated einy changes in the Lamp from one phase of the study to 
another. The UV system was built so that the lamp could be 
quick-ly moved without being shut off from one water layer to the 
next to minimize changes in the effluent and flow rates. 

Phase 2 showed that the design flow rate for the raw effluent 
was 241 litres per minute and for the primary effluent it was 522 
litres per minute. This was equivalent to a UV dose of 13mW. 
sec/itiL for the raw effluent and 6mW. sec/mL for the primary 

3. Materials ani3 Methods 

A UV unit (Figure 16) was built with three different water 
layers and a movable 2000 watt medium pressure mercury lamp. 

Raw or primary effluent was pumped from the Greenway 
Wastewater Treatment Plant in London. Ontario, Canada through the 
UV unit at a the flow rates shown in Figures n-28. These flow 
rates varied from 110 L/min to 796 L/rain. The average flow rate 
was kept as close as possible to the optimum flow rate shown in 
Table 6 for raw and primary effluent. The flow rate was measured 
with a stopwatch and 400 litre reservoir. 

The UV unit was operated in a vertical position and the 
effluent was pumped Into the bottom of the system. After the 
initial minimum flow rate was set, Che lamp was turned on and 
allowed to burn until the amperage of the lamp stabilized. 

The lamp was moved by pulling a marked wire which protruded 
from top and bottom of the UV unit. The marked wire positioned 
the UV lamp in Che middle of each water layer. 





- — PIPE OD 16H lO 163 

PIPE CO 114 lO MO 

PIP£ OO 76 lO 73 



Figure 16: A schematic diagram of the continuous flow IIV unit with three different water 
Layers and a noveable UV lamp. 

At the beginning of each day's run, a 3.25 kg bottle of 
analytical reagent grade nitric or hydrochloric acid (British Drug 
House, Toronto, Canada} was used to clean the quartz sleeve. The 
dcid and water were poured into the UV unit until the quartz 
sleeve was submerged and then the UV system was gently rocked to 
mix Che acid and the water. A visual inspection showed that this 
procedure cleaned the quartz sleeve. 

Each water layer was tested at one flow rate and then the UV 
lamp was turned off and the flow rate changed. 

Two samples were collected at each flow rate for every water 
layer. One was immediately put on ice in the dark amd the second 
sample was sxibjected Co sunlight according to the method of Whitby 
ec al. (1984) . 

Each sample was analyzed for UV cransnictance at a wavelength 
o£ 254 nm. 

The total suspended solids of each wastewater was analyzed 
according to Method 209C In the 16th Edition of Standard Methods 
for the Examination of Water and Wastewater (American Public 
Health Association, 1985). 

The fecal colifonns were measured by the membrane filtration 
method {Ontario Ministry of the Environment, 1984) . 

4 . Results and Discussion 

a- Raw Effluent 

The level and fraction survival of the fecal coliforms in the 
raw effluent after irradiation with the 2000 watt medium pressure 
mercury lamps, three different water layers and various flow rates 
are shown in Figures 17-22. Each Figure shows the level or 
fraction survival of che fecal coliforms before and after three 
hours in the sunlight. 



: Control 



/ ' 


/ • 


.• A/ 









* / 





'/ a • 






' • 







*• " / 


* y 









100 200 300 



Fi^re 17: The effect of Che tw cencimecre water layer of Che continuous flow 
UV reactor on the number of fecil conforms In raw effluent after 
(A) and before (Bl phocoreactlvacion. 








50 100 150 200 250 300 350 ^00 


Figure 18: The effect of che Cud cenCLnecre water layer of Che continuous flow 
UV reactor on che fraction survival of che fecal collforins In raw 
effluent afcer (A) and before (B) phocoreacctvaClon. 


10 '^:t Control 








< 10' 



100 200 300 



Figure 19: The effect of the 3.85 centimecre water layer o£ Che continuous 
tLou UV reaccor on Che number of fecal colLforms tn raw effluent 
afcer (A) arxl before (B) phocoreaccivacion. 








50 100 150 200 250 300 350 


Figure 20: 

The «££ecc ai the 3.85 centimetre water layer of the continucxjs 
flow UV reactor on the fraction survivaL of the fecal coliforms 
in raw effluent after (A) and before IB) phocoreactlvatton. 

108 n 

10^:: Control 








100 200 300 



Figure 21; 

The effect of ch* 6-5 centl«tre waCeT layer of the continuous 

ISr^At^^^Lr '^pV^' °^ ^^"^ coUforms in raw effluent 
atcer (A) and before (B) photoreaccivatton. 




01 10' 



50 100 150 200 250 300 350 


Figure 22; The effect of Che 6.5 centimecre water layer of Che continuous 
fl»j UV reacCor on the fraction survival of che fecal collforms 
in raw effluent afcer (A) and before (B) photoreaccivacion. 

The unflltered raw effluent had a percent transmifision of 
12.7 (SD=1.7) at a wavelength of 254 nm. The level of suspended 
solids in the raw effluent was 179 mg/L (SD=41.2). 

The dose of UV light in the continuous flow reactor can be 
calculated by dividing the germicidal wattage of the lamp by the 
flow rate in millilitres per second. 

The total germicidal output of the 2000 w medium pressure 
lamp was 52.2 watts as measured in Phase 2. Ten percent of this 
output was subtracted for loses through the quartz sleeve. The 
dose of UV light In the continuous flow reactor at the slowest 
flow rate (110 L/min) was 26mW. sec/mL and 7mW. sec/mL at the 
highest flow rate (399 L/min) . The experiments with the 
colllmated beam showed that a dose of 13mW. sec/mL was required 
for a three logarithm reduction of the fecal conforms without 
photoreactivation . 

When the above doses of UV light are compared with those in 
the experiments with the colllmated beam (Phase 2}. the continuous 
flow reactor was unable to reduce the fecal coliforms to the same 
concentration or reach the same fraction survival. 

The dry summer in London, Ontario, Canada had increased the 
suspended solids by 46 percent and decreased the transmission of 
the UV light by 54 percent. The increased concentration of 
suspended solids shields more fecal conforms from the UV 
radiation and thus the limiting number of microorganisras which can 
be killed increases. 

Mixing of Che effluent is more complete during the 
experiments with the colllmated beam and this will result in a 
greater kill of the fecal coliforms when the same volume of fluid 
is subjected to an identical dose of UV light. 


A three logarichm kill of the fecal conforms waB obtained 
with flows o£ less than 172 litres per minute with the two 
centimeter water layer and 2000 watt medium pressure lamp. After 
photo re activation, a three logarithm kill of the fecal coliforms 
was not obtained at any of the tested flow rates. Extrapolating 
the data beyond the tested flow rates may be invalid because the 
dose response curve is not linear over the entire range of UV 
doses. This is Illustrated by the dose reaponse curves in Phase 1 
(Figures 1-8). If the response is linear, then the two centimeter 
water layer produces a three logarithm kill after 

photoreactivation at a flow rate of 14 litres per minute and the 
water layer with a depth of 3.85 cm produces a three logarithm 
kill without photoreactivation at a flow rate of 11 litres per 

As the kill of the fecal coliforms in the raw effluent 
increased there was a general increase in the degree of 
photoreactivation and this is in agreement with the research 
reviewed by Rupert (1964). 

b. Primary Effluent 

The level and fraction survival of the fecal coliforms in the 

primary effluent after irradiation with the 2000 watt medium 

pressure mercury lamp at various flow rates through three 

different water layers are shown in figures 23-28. Each Figure 

shows the level or fraction survival of the fecal coliforms before 
and after three hours in the sunlight. 

The unfiltered primary effluent had a percent transmission of 
27.1 (SD=1.7) at a wavelength of 254 nm. The level of suspended 
solids in the primary effluent was 45 mg/L (SD=6.7) . The percent 
transmission was almost identical to that during the experiments 
with the collimated beam. The level of suspended solids was 27 
percent less during the continuous flow studies. 


10^ i Control 

10'' ■ 








100 200 300 -^.OO 500 600 700 800 

Figure 23: The effect o£ Che cms cenclmecre uacer layer of che concinuous flow 
W reactor on che number of fecal coUforms in primary effluent 
after (A) and before (B) [rfiotoreaccivBClon. 

100 200 300 ^00 500 600 700 800 

Figure 24; The effecc of Che two centimcCres water layer of Che conCinuous 
flow UV reactor on the fraction survival of the fecal coUforms 
in prlMry effluent after (A) and before (B) photoreacCivatlon. 

10*^ i Control 








100 200 300 ^00 500 600 700 800 

Figure 25: The effect of the 3.85 cenctmeCre water Layer of Che continuous 
flow W reactor on Che rajmber of fecal collforms in primary 
effluent after (Al and before (B) photo reactivation. 

100 200 300 ^00 500 600 700 800 

Figure 26: The effect of the 3.85 cenclinecre water layer of the continuous flow 
UV reactor on the fraction survival of the fecal colifonrs in 
primary effluent after (A) and before (B) photoreacclvatlon. 

lO"^ ^Control 




100 200 300 ^00 500 600 700 


Fi^re 27: The effect of Che 6.S c«ncimecre wacer layer of Che continuous flow 
UV reaccor on the number of fecal coltfoms In primary effluent 
after lA) and before (Bl ;^To:oreacclvacton. 


5 10-1 


g 10-2 



5 10"'^ 







100 200 300 AOO 500 600 700 800 

Figure 28: The effect of che 6.5 cencimetre wacer layer of Che conCinuous flow 
UV reactor on che fracclon survival of Che fecal coliforms in 
primary effluenc after (A) and before (B) phocoreactivation. 

The dose of UV light in the continuous flow reactor was 
calculated in the same manner as it was for the raw effluent. The 
dose of UV light in the continuous flow reactor at the minimum 
flow rate (113 L/min) was 25 mw. sec/mL and 3.5 mW. sec/mL at the 
maximum flow rate (796 L/min) . The experiments with the 
collimated beam in Phase 1 showed that a dose of 6 raW, sec/mL 
produced a three logarithm kill of the fecal coliforms without 
photoreac t i va t i on . 

A comparison of the UV dose response between the continuous 
flow reactor and the experiments with the collimated beam showed 
that the two centimeter water layer attained the same or a better 
kill of the fecal coliforms at the high and low flow rates. 
Because the two sets of primary effluents were more closely 
related than the raw effluents, the mixing occurring during the 
Clow of water through the two centimeter water layer must be 
similar to that occurring in the dishes used during the 
exi>eriments with the collimated beam. 

The 38.5 mm water layer also attained a three logarithm kill 
of the fecal coliforms before photoreac tivat ion at the lowest flow 
rate which was tested. None of the combinations of flow rate and 
water layer produced a three logarithm kill of the fecal 

The data from the 65 mm water layer can be extrapolated to 
obtain the flow rate which produces a three logarithm kill of the 
fecal coliforms. This flow rate should be used with caution 
because the dose response curve is not linear over the entire UV 
dose range as shown in Figures 2-8. Without photoreactivation, 
the flow rate was 34 litres per minute Cor the 65 mm water layer. 

Photoreactivation resulted in an average 1.3 logarithm 
increase in the fraction survival of the fecal collCorms in 
primary effluent. The average increase due to photoreactivation of 
the fecal coliforms in the raw and primary effluent after UV 
irradiation was one logarithm. This is similar to other studies 


with UV irradiated fecal coli forms in wastewater (Whitby et al. 
1985 and 1984; Scheible and Bassell, 1981 and Bohm et al., 1982). 
There was no consistent increase or decrease in the degree of 
pho tore activation of the fecal coliforms in Che primary effluent 
with a change in flow rate or water layer. 


1. The two centimeter water layer with the 2000 W medium 
pressure mercury lamp approached the kill of the fecal 
coliforms obtained with the experiments with the collimated 
beam in Phase 1 . 

2. Mixing of effluents with high concentrations of suspended 
solids and low UV transmission is essential because each 
microbe must receive a minimum dose of UV light to be 
destroyed. Only the two centimeter water layer approached 
the proper mixing regime with raw and primary effluent. 

3. A dose response curve of the fecal coliforms in low quality 
wastewaters should be prepared using static and continuous 
flow methods. The differences between the effluents and the 
results of the experiments with the collimated beam and the 
continuous flow reactor show the need for these types of 

4. The UV equipment must be built for the worst conditions 
because a change in the quality of the effluent can 
dramatically effect the performance of the UV unit. 

5. The continuous flow reactor was able to reach a three 
logarithm kill of the fecal coliforms with and without 
photoreactivation when treating primary effluent but only 
without photoreactivation when treating raw effluent. 




1. Introduction 

An analysis ot capital, operating and maintenance costs of 
disinlecting primary and raw effluents with mediuni pressure 
mercury lamps was undertaken to compare this disinfection 
alternative with chlorinatlon and UV irradiation with low pressure 
mercury lamps, 

2. Design Specification 

a. Low Quality Wastewaters 

Very little information is available which describes the UV 
transmission or concentration o£ suspended solids in low quality 
wastewaters. The simulated combined sewer overflow used by Zu)tovs 
et al. (1986) had a UV transmission at a wavelength of 254 nm o£ 
2.B percent per centimeter pathiength and Che level of suspended 
solids was 187 mg/L. The primary effluent studied by Scheible 
et al. (19S5) had a UV transmission at a wavelength of 254 nm of 
55 percent per centimeter and the concentration of suspended 
solids was 80 mg/L. A review of the literature by Zukovs et al. 
(1986) showed that combined sewer overflow had levels of suspended 
solids between 80 and 274 rag/L. 


Due to the above variations in low quality wastewaters, costs 
were developed for raw and primary effluent with the following UV 
transmissions at a wavelength of 254 nm and level of suspended 
solids. The raw effluent had a minimum UV transmission at a 
wavelength of 254 nm of 13 percent and a maximum concentration of 
suspended solids of 180 mg/L. The primary effluent had a minimum 
UV transmission at a wavelength of 254 nm of 27 percent and a 
maximum concentration of suspended solids of 45 mg/L. 

b. Disinfection Standard 

Phase 1 of this project showed that it was not practical to 
strive for a four logarithm kill of Che fecal conforms. Kollar 
et al. (1986} estimated their UV irradiation and chlorination 
costs with a four logarithm reduction of fecal conforms but the 
results in this study show that this Is not possible with UV 

A three logarithm kill of the fecal coliforms was obtained in 
the experiments with the collimated beam (Phase 1) and the 
continuous flow reactor (Phase 3) . The UV equipment was costed to 
reach a three logarithm kill of the fecal coliforms. 

If a three logarithm kill of the fecal conforms after 
photoreactivation is required then it Is not practical to treat 
raw effluent but it is possible to treat primary effluent. In raw 
effluent none of the combinations of flow rate and water layer 
reached a three logarithm kill of the fecal coliforms after 
phocoreactlvacion. In primary effluent a three logarithm kill of 
the fecal coliforms was obtained after photoreactivation. 

3. Disinfection Facilities 

a. Introduction 

UV equipment was designed for three different peak flow 
rates: 5,000, 50,000 and 500,000 mVday. The UV systems were 
designed for raw effluent without photoreactivation of the three 
logarithm kill of the fecal coliforms and tor primary effluent 
with a three logarithm kill of the fecal coliforms before and 
after photoreactivation. 


This cost analysis was £or the capital, operating and 
maintenance cost for the UV equipment only and not the flow 
meters, buildings, cement work. etc. 

b. Raw Effluent 

The basic unit for estimating capital costs was a medium 
pressure mercury lamp and its associated hardware such a quartz 

sheath, ballast, starter, lamp supports, control box, cooling 
equipment and stainless steel shell. The capital costs tor the UV 
equipment are shown in Table 7, 

The operating and maintenance coses are shown in Table l. 
The lamp replacement costs were estimated to be S300.00 per UV 
lamp. The UV lamps must be replaced every 5000 hours. Kollar et 
al. (1986J assumed that these facilities would operate only when 
required during the period from March to November, with total 
annual operating time assumed to be 250 hours. The lamp 
replacement frequency was assumed to be once every twenty years 
for the medium pressure mercury lamps. 

After each use, the quartz sheaths should be acid washed to 
remove any deposits. A ten percent solution of phosphoric acid 
can be recirculated through the UV unit. This solution can be 
stored for further use. The estimated time required for cleaning 
was 4.5 to 8 hours. The capital cost for the inplace deeming 
systems is shown in Table 8. The wage rate for a worker for the 
Ontario Ministry oE the Environment is 313-41 per hour plus 25 
percent benefits. 

The power costs are shown in Table 7. 


Table 7: Esclmaced coscs for UV disinfection of raw effluerc wlchouc 
phocoreaccivation of the fecal collEorms 

Cost Componenr 
Capital UV SysCem 

In Place Cleaning 

local Capical (S 1988) 
Operation and Maintenance 

Power (6(/Io*i) 

Ubour (S16-76/h} 



Total Operation and Maintenance 
(S/year 198S) 

flow Rate (n^/dl 

5,000 50,000 500,000 

96,000 768,000 7,680,000 

14,000 22.000 150.000 

110,000 790.000 7,630,000 

Z30i 23,040 230,«» 

1676 2,095 3,3» 

240 2,AO0 24,000 

24 241 2,410 




c. Primary Effluent 

The two centimeter water layer was able to produce a three 
logarithm kill of the fecal collforms with and without 
photoreactivation at flow rates of 360 and 825 litres per minute 
per UV lamp. The 3.B5 centimeter water layer was able to produce 
a three logarithm kill fo the fecal collforms without 
photoreactivation at a flow rate of 520 litres per minute per UV 
lamp. Extrapolation of the data with the 3.85 centimeter water 
layer showed that a three logarithm kill of the fecal coliforras 
after photoreactivation could be obtained at a flow rate of 175 
litres per minute per UV lamp. 

A flow rate of 370 and 147 litres per minute per UV lamp was 
used for estimating the number of lamps for a three logarithm kill 
of the fecal collforms before and after photoreactivation, 
respectively. This Is the average flow rate between the 2 and 
3.85 centimeter water layer decreased by 45 percent for lamp 

The capital costs for the UV equipment are shown in Tables 8 
and 9. 

The operating and maintenance costs for primary effluent were 
estimated in an identical fashion to that of the raw effluent and 
are shown in Tables 8 and 9. 

The annualized use-costs for UV disinfection of raw and 
primary effluent presented in Table 10 are based on a 20 year 
period and a discount race of 1% as was used by Kollar et al. 
(1986) . 


Table 8: Estimated cosCs for UV disinfection oE primary effluent without 
phocoreactivaCion of Che fecal colifonns 

Flow Race (m /d) 

Cost Componenc 5.000 50,000 500,000 

Capital UV System 18,000 l-W.OOO 1,«0,000 

In Place Cleaning 7,000 11.000 75.000 

Total Capital !t 1988) 25,000 155.000 1,515,000 

Operation and Maintenance 

Power (6#/ta*) 

Labour l$l6.76/h! 



Total Operation and Malncenance 2,158 b,^98 50,315 

1 I/year 1988) 













Table 9: Escimaced cos:s for W dlslnfecclon of primary effluent: wich 
phoCore activation of the fecal colifonns 

Cost Conponenc 
Capital UV SysCOQ 

In Place Cleaning 
Total Capital ($ 1988) 
Cperatlon and Maintenance 

Power (64/kuh) 

Ubour (Sl6.76/h) 



Total Operation and Maintenance 
($/year 1988} 





Flow Rate (m-'/d) 

5,000 50,000 500,000 

36,000 288,000 2,880,000 

14,000 22,000 150.000 

50,000 310,000 3,030.000 

8.640 86,400 

1,676 2,095 

900 9,000 






Table 10: EsClmaCed Use-Cosc of Disinfecting Prtnary and Raw Effluents with 
Medium Pressure Mercury Lairps 

DjsC Coaiponenc Flowrate (n 16) 

($1988) 5000 50,000 500,000 

Raw, No PhocorcacClvaclon 
Prlaary, No PhoCoreactlvaClon 

Primary, Phocoreaccivation 










The escimated use-costs of disinfection simulated combined 
sewer overflow and chemically treated primary effluent with low 
pressure mercury lamps are shown in Table 11. These costs were 
from the study of Kollar et al. (1986). To compare the two types 
of UV lamps only the capital and operating costs of the UV 
equipment itself were considered. 

To compare UV disinfection with chlorination/dechlorination 
only the contact chamber, chemical storage tanlts, pumps, piping, 
flow meter, Injector system, evaporator, operation and maintenance 
were considered in the use~cost estimate from the work of Kollar 
et al. (1986) and these use-costs are shown in Table 12. The 
majority of the other capital costs are common to all of the forms 
of disinfection. 

Disinfection of low quality wastewaters with UV light from 
medium pressure mercury lamps or with chlorinatlon is lower than 
with low pressure mercury lamps. 


Table 11: Estlmaced Use-Cost of DlslnfccClng Simulaced Combined Sewer 

Overflow and Chemically Treated PrLnary EfflgenC with Low 
Pressure Mercury Lamps 

Cose Cofflponent Flourace (ni-^/d) 

(S1985I 5000 M.OOO 500,000 

Simulated Combined IS.gS/rr-'/d/yr 12. W 12.04 
Sewer Overflow 

Chemically Treated 11.70 10.87 10.12 
Priiftar)- Effluent 

Table 12: Estlmaced Use-Cosc of DlsinfecCing Simulated Combined Sewer 

Overflow and Chemically Treated Primary Effluent with 
Chlorinacion/Dechlorlnatlon and Chlorination, Respectively 

Cost Component Flowrate (tn /d) 

((1985) son 50,000 500,000 

Simulated Combined ■, 

Sewer 0>«rf low 2.48/»-*/d/yr 1.36 l.<» 

Chemically Treated 

Primary Effluent 1.26 0.37 OvW 


1. Each type of wastewater retjuired a ciifferent dose of UV light 
to reach the required level of disinfection due to the UV 
transmission, suspended solids and the relationship of the 
fecal conforms with the solids. 

2. To design a UV system for low iiuality wastewaters a series of 
survival curves should be prepared using the method with a 
colllmated beam to determine whether the disinfection 
standard can be attained and the proper dose of UV light 
which is required to reach this disinfection standard. These 
results should be confirmed with continuous flow studies with 
a scale model of the projected UV equipment. 

3. Medium pressure mercury lamps can reduce the fecal coliforms 
in raw and primary effluent by three logarithms in a static 
and continuous flow situation, 

4. A use-cost comparison of UV disinfection of raw and primary 
effluent with medium pressure mercury lamps with that with 
low pressure mercury lamps showed that capital, operating and 
maintenance costs were lower with the former lamps. 

5. Treatment of primary effluent by medium pressure mercury 
lamps without photoreactivation of the fecal coliforms was 
use-cost competitive with the chlortnation of chemically 
treated primary wastewater, Chlorination and 
chlorinotion/dechlorination were lower in cost for the other 

6. UV disinfection of low quality wastewaters may be an 
alternative to chlorination when the ecological 
considerations are taken into account such as the production 
of harmful chloro-organic compounds and residual of chlorine 
on the aquatic biota. 



American Public Health Association. 1985. Steuidard Methods for 
the ExBinlnstion of Water and wastewater. 16th ed. American 
Public Health Association, Washington, D.C. 

Bohm. P., K.W.A., Ho and J,E. Pagel. 1982. Application of UV 
Disinfection Technology In Ontario Water Pollution Control Plant 
Effluents. Ontario Ministry of the Environment Technology 
Transfer Conference No. 3. Toronto. Ontario, Canada, December 7. 

Cochrane Division, Crane Co. 1970, Microstraining and 
Disinfection of Combined Sewer Overflows.. Water Pollution 
Control Research Series 11023 EVO 06/70. U.S. Department of The 
Interior, Washington D.C, U.S.A. 

Drewhing, F.J., Oliver, D.A. MacArthur and P.E. Moffa. 1979. 
Disinfection/Treatment of Combined Sewer Overlfows, Syracuse, New 
York. EPA-600/2-79-134. U.S. Environmental Protection Agency, 
Cincinnati, Ohio, U.S.A. 

Dutka, B.J. and S- Tobin. 1976. Monitoring of Storm Water 
Runoffs for Bacterial and Viral Pathogens of Han. Project No. 
76-8-41, In: Microbiological Characteristics of Urban Storm 
Water Runoffs in Central Ontario, Research Report No. 87. 
Canada-Ontario Agreement on Great LaJtes Water Quality. Ontario 
Ministry of the Environment, Toronto, Ontario, Canada. 

Ho, K.w.A. and P, Bohm. 1961. UV Disinfection of Tertiary and 
Secondary Effluents. Water Pollution Research Journal of Canada 
16: 33-44. 

Kollar, J., G. Zukovs. H.D. Monteith. 1986. An Assessment of 
Ultraviolet Irradiation for Disinfection of Low Quality 
Wastewater, Project 83-304 for the Ontario Ministry of the 
Envi ronmen t . Toronto , Canada . 


HacLaren, J. P. 1980. Stormwater Management Technology Systems 
Demonstration in the City of St. Thomas. Report SCAT - 3. 
Information and Communications Centre, Canada Mortgage and 
Housing Corporation. Ottawa, Ontario, Canada. 

Oliver. B.C. and E.G. Cosgrove. 1975. The DisinCection of Sewage 
Treatment Plant Effluents Using Ultraviolet Light. The Canadian 
Journal o£ Chemical Engineering 53::70-n4 (April 1975). 

Ontario Ministry of the Environment. 1984. Handbook o£ 
Analytical Methods for Environmental samples. Ontario Ministry of 
the Environment, Toronto. Ontario, Canada. 

Quails. R.G. and J.D. Johnson. 1983. Bioassay and Dose 
Measurement in UV Disinfection. Applied and Environmental 
Microbiology 45: 872-877. 

Quails, R.G., S.F. Ossoff, J.C.H. Chang, M.H. Dorfman, CM. 
Dumais, D.C. Lobe and J.D. Johnson. 19B5. Factors Controlling 
Sensitivity in Ultraviolet Disinfection of Secondary Effluents. 
Journal Water Pollution Control Federation 57: 1006-1011. 

Qureshi. A. A. 1974. Microbiological Characteristics of storm 
Water Runoffs at East York (Toronto) and Guelph Separate storm 
Sewers. Project No, 74-8-25. In: Microbiological 
Characteristics of Urban Storm Water Runoffs in Central Canada, 
Research Report No. 87. Canada -Ontario Agreement on Great Lakes 
Water Quality. Ontario Ministry of the Environment, Toronto, 
Ontario, Canada. 

Rupert. C.S. 1964. Photoreactivation of Ultraviolet Damage, in 
Photophysiology volume 2. Edited by A.C. Giese, Academic Press, 
New York, U.S.A. Pages 283-327. 


Schelble, O.K. and CD. Bassell. 1981. Ultraviolet Disinfection 
or a Secondary Wastewater Treatment Plant Effluent. 
EPA-600/2-81-152, U.S. Environmental Protection Agency, 

Cincinnati, Ohio, U.S.A. 

Scheible, O.K. M.C. Casey and A. Forndran. 1985. Ultraviolet 
Disinfection of Wastewaters from Secondary Effluent and Combined 
Sewer Overflows. EP/600/2 -66/005. U.S. Environmental Protection 
Agency. Cincinnati, Ohio, U.S.A. 

U.S. Environmental Protection Agency, 1986. Design Manual, 
Municipal Wastewater Disinfection. EPA/625/1-86/021. U.S. 
Environmental Protection Agency, Cincinnati. Ohio, U.S.A. 

Waller, D.H. and Z. Novak. 1975. Municipal Pollutant Loadings to 
the Great LaJtes from Ontario Communities, Research Report No. 94. 
Canada-Ontario Agreement on Great Lakes Water Quality, Ontario 
Ministry of the Environment. Toronto, Ontario, Canada. 

Whitby, G.E., G. Palmateer, W.G. Cook, F. Soon and E. Janzen, 

1985. The Effects of Wastewater Quality on Ultraviolet Light 
Disinfection. Ontario Ministry of the Environment. Technology 
Transfer Conference No. 5. Toronto, Ontario Canada. 

Whitby , G . E . , G . Palmateer . W.G. Cook, J . Haarschalkerweerd . D . 
Huber and K. Flood. 1984. Ultraviolet Disinfection of secondary 
Effluent. Journal Water Pollution Control Federation 56 : 

Zukovs. G., J. Kollar. H.D. Monteith. K.W.A. Ho and S.A. Ross. 

1986. Disinfection of Low Quality Wastewaters by Ultraviolet 
Light Irradiation. Journal Water Pollution Control Federation 58: 






P.L. Seyfried, T. Bleier, Y. Xu and R. Harmandayan, 
Department of Microbiology, FitzGerald Building, 

University of Toronto, Toronto, Ontario 

This study, sponsored by the Ministry of the Environment, 
investigated the use of specific bacteria to detect human fecal 
wastes in storm sewer lines. The organisms examined were fecal 
col i forms , Escherichia coli , fecal streptococci , enterococci , 

PS^WdQinpngg aeruginosa . Clogtridiuro perfrinqens . and 

Bif idobacter4i;]n sp. These bacteria were isolated during periods 
of wet and dry weather from surface runoff, from designated 
locations in sanitary sewer lines, and from priority and non- 
priority storm sewers. Biochemical testing, serotyplng, and/or 
genotyping were used to further characterize more than 4 , 000 
fecal streptococcus, Pseudomonas aeruq^ngafl . and Bifidobacterium 
isolates. Speciatlon of the fecal streptococci showed that 

Streptococcus faeca;j.g subsp. faecalip was more predominant in 

sanitary and high priority sewers than in surface runoff and 
non-priority sewers. S. casseliflavMs , on the other hand, was 
primarily found in runoff and non-priority storm sewers. DKA 
sequence studies of the fecal streptococci, using Restriction 
Endonuclease Analysis (REA) produced many different patterns and 
it was difficult to establish any relationship between the 
isolates. By comparison, P. aeruginosa genotypes were more 
uniform and fewer patterns were observed. As an example, a 
specific P. aeruginosa genotype was isolated from both the street 
runoff and the storm sewer at one City of Toronto location. 
Genotyping also appears to be a good method of distinguishing 
between the Bif idqbacterium sp. that are predominant in sewage. 



The purpose of storm sewers Is to collect storo water from 
urban areas and channel it into receiving waters such as rivers, 
streams, or lakes. Because storm sewer water does not receive any 
treatment prior to its discharge, it is important to identify any 
illegal sanitary connections in the storm sewer line. A small 
aaount of dry weather flow would be expected in a normal or non- 
priority storm sewer; however, a priority storm sewer outfall 
that discharges more than I L/sec during dry weather and contains 
fecal coliform (FC) levels greater than 10,000 FC/lOO mL is 
highly suspect and should be investigated. Because a method was 
needed to identify the source of human fecal pollution in storm 
sewers, a study characterizing the bacterial indicators found in 
both sanitary and storm sewers was initiated in the fall of 1986. 

In the first year of the study, samples were collected from 
surface runoff when it rained and from specific locations in 
sanitary sewer lines and in priority and non-priority storm 
sewers during periods of wet and dry weather. Fecal coliforms, 
Escherichia coli . fecal streptococci , enterococci , Pseudomonas 
aeruqjnpga . Clostridium perfrinoens and Bif idobacteriura sp. were 
enumerated in each of the samples. The results, presented in 
Part D of the 1987 Technology Transfer Conference Proceedings 
(Sey fried s^ al. . 1987) showed that higher densities of all 
indicator organisms, including Bjfidobjicterium . were recovered 
from priority stom sewer samples than from non-priority sewage 


samples. It was found that chancfes in bacterial Indicator 
populations were best observed during periods of dry weather. 

The objective of the segment of the study described herein 
was to analyze more dry weather samples and further characterize, 
by means of biochemical testing, serotyping and/or genotyping, 
the fecal streptococci, Pseudomonas aeruginosa , and 
Bifidobacterium isolates from sanitary and stom sewers. 

Sampling sites 

Sites A, B and C (shown in Table 1 and Fig. l) in the Mount 
Steven Trunk storm sewer line were sampled because this area was 
designated a high priority sewer by the Ministry of the 
Environment. The non-priority sites, selected for comparison, 
were X, Y and Z in the Mount Steven Trunk storm sewer branch 
lines. During periods of wet weather, storm water run-off was 
also collected at the X, Y and 2 sites. These samples were 
labelled R, G and Q, respectively. Samples D, E and F were 
obtained from a sanitary sewer in close proximity to the priority 
storm sewer sampling points. 

Sample Collection 

During periods of dry weather, triplicate samples were 
collected from each sampling point in the sewers over a four-day 
period. The dry weather samples were obtained in October, 1986; 
June, 1987; August, 1987; June, 1988; and August, 1988. Wet 


TABLE 1. Sampling locations of high priority and non-priority 
storm sewers, sanitary sewer and stom water runoff. 




High Priority 
storm Sewer Line, 
Mount Steven 
Stonti Sewer Trunk 

Danforth and Jones Avenues - furthest 
in-line sampling point [near source of 
suspected solution input). 

Pape and Strathcona Avenues (mid-line 
sampling point) , 

First and Broadview Avenues (near outfall}. 

Storm Sewer 
Branch Lines 

Chatham and Jones Avenues (connects to main 
line above sampling point B) . 

Danforth And woodycrest Avenues (connects 
to main line above sampling point A). 

Pape and Cavell Avenues. 

Storm Water 

Chatham and Jones 

Danforth and Woodycrest Avenues 

Pape and Cavell Avenues 

Sewage Line 

Danforth and Jones Avenues 
Strathcona and Pape Avenues 
First and Broadview Avenues 


z , ,(Q) 


^ ♦^ 




F!G. 1 - Schematic diagram of Mount Steven storm sewer and sanitary sewer lines. 


weather samples were collected from the sewers and the street 
run-off during rainy days in July, September and October, 1987. 

The samples were collected in sterile glass containers, 
transported to the laboratory on ice, and processed within 6 
hours of collection. 

Fecal specimens from both humans and animals were also 

Bacterial Isolation and Enn ip.e r fl t . i.g n 

Samples were analyzed for fecal col i forms using m-TEC agar 
{Dufour, 1981), Escherichia coli by means of the m-TEC urease 
treatment described by Dufour (1975; 1981) , fecal streptococci 
isolated on m-Enterococcus agar (Slanetz and Bart ley, 1977) , 
enterococoi on m-HE agar (Dufour, 1980) , Pseudomonap a^yuginoga 
using m-PA agar (Standard Methods, 1985) and Bifidobacterium sp. 
isolated on the VN-17 medium described by Mara and Oragui (1983). 

All bacteria were identified to the species level using 
standard taiconomic methods (Seyfried et_sUj.i 19S7). 

Bacterial Characterizatiop 

Approximately 2,500 fecal streptococci were recovered on m- 
EnterococGUs agar and m-ME agar during the dry and wet weather 
surveys. The isolates that were identified as varieties of S. 
faecal^p were tested for their reaction in litmus milk broth 
(Difco) using the method of Mundt (1973). 

Serotyping of the P. j ieruginosa isolates was carried out 
using a Pseudomonas Antisera Kit (Difco) . The organisms were sub- 


speclated into the 17 different heat-stable somatic antigen 
groups described by Kusama (1978). 

For the restriction enzyme analysis (REA) of P. aeruginosa , 
total cellular DNA was extracted using a method described by 
Bradbury et al. (1984, 1985) . A 1.5 mL volume of an 18 hour 
nutrient broth culture inoculated with P. aeruginosa was 
transferred into an Eppendorf tube and centrifuged in a Microfuge 
12 (Beckman) for 3 minutes at 7500 x g. The supernatant was 
discarded and the pellet loosened by vortexing. A 291 ;jL volume 
of PEE 1 buffer containing 10 rng/mL lysozyme was added and the 
mixture incubated for 20 minutes at 35* C. A 9 tiL amount of 5H 
NaCl was added, thoroughly mixed, and then 150 pL of 10% SDS was 
added. The solution was gently mixed and incubated for 10 minutes 
at 37* C. Following the addition of 450 pL 
phenol : chloroform : isoamyl (25:24:1), the mixture was vortexed 
and centrifuged at 7500 x g for six minutes at room temperature. 
The upper aqueous phase was removed with a pasteur pipette and 
transferred to an Eppendorf tube. One mL of 95% cold ethanol was 
added, the tubes vigorously shaken and stored at -20' C 
overnight. The mixture was centrifuged for 3 minutes at 12, 000 
X g, the supernatant discarded and the pellet redissolved in 250 
mL DNA wash buffer. A total of 500 ^lL. of 95% cold ethanol was 
added, the mixture stored at -20* c for 20 minutes, and 
centrifuged at l , 200 x g for 3 minutes . The supernatant was 
discarded and the pellet allowed to dry at 37* C for 10 minutes. 
The pellet was dissolved in 100 yL of distilled water and stored 


at 4 " c until digested. 

Restriction digests were performed using Sma I according to 
the manufacturer's instructions (Boehringer Mannheim). A 10 yh 
aliquot of double strength {2X) Sma I buffer was placed in an 
Eppendorf tube and 10 uL extracted DMA added. A 2 >]L sample of 
Sma I enzyme was added and the mixture was incubated for 1 hour 
at 37" C to allow for complete digestion. Following the addition 
of 1 yL of 0.15M EDTA + 0.4 ng/mL RNase A, the tube was incubated 
at 37" c for 20 minutes, A 5 jiL volume of 5X sample buffer was 
then added to the restriction digest. Samples were 
electrophoresed on 0.7% agarose gel for 16 hours at 27 volts. 
Gels were stained with 1 mg/mL ethidium bromide in IX TAE (Tris 
base, 1.0 sodium acetate, 0.1 M disodium EDTA) for 1 hour and 
destained for 2 hours in distilled water. Photography was done 
using U.v. light at 300 nm and a red No. 23A polaroid 665p/N film 
with an exposure time of 30 seconds. 

Fecal streptococci were grown in Brain Heart Infusion broth 
(Difco) at 37' C for 4 hours and the total cellular DNA extracted 
as described previously. Restriction digests were performed using 
Bam HI enzyme according to the manufacturer's directions 
(Boehringer Hannheim) along with a 10 uL aliquot of 2X Bam HI 

Bj-f j-dobacterium sp. were grown anaerobically in MRS broth 
(Oxoid) for 48 hours at 37" C. Three different enzymes, Sma I, 
Bam HI and Cfo, with their appropriate buffers, were used for the 
restriction enzyme analysis of the bifidobacteria. 



Data from the previous study (Sey fried et al . . 1987) 
suggested that fecal contamination, possibly human in origin, was 
evident in the storm sewer line near sites A and V. The results 
of the 1988 survey, presented in Table 2, add support to this 
conclusion. As may be seen in the table, the fecal col i form 
levels in the high-priority storm sewer were greater than 
10,000/100 mL at site A in June and at all sites in August. It 
should be noted that counts of all indicator organisms tended to 
be higher in August, possibly due to regrowth of the bacteria tn 
the warmer nutrient-enriched waters (Hoadley, 1977). 

As might be expected, fecal coliform and fecal streptococcus 
counts were highest in sanitary sewage and the fecal coliform to 
fecal streptococcus ratio was greater than 4 in these samples. 
Although a ratio greater than 4 was observed at site A in the 
priority storm sewer, at other storm sewer sites the FC/FS ratios 
were generally below -l indicating that there was little or no 
human fecal input (Geldriech and Kenner, 1969). 

P. aeruginosa and Bif l<4Qt?t>cterium sp. were shown in the 1937 
study to have potential as indicators of human fecal waste. 
Collaborative data presented in Table 2 shows that counts of both 
organisms were higher in sanitary and priority storm sewage than 
in non-priority storm sewage. 

Hara and Oragui first proposed the use of sorbitol 
fermenting bifidobacteria as indicators of human fecal pollution 


TABLE 2. overall qBiMKCrlc mean conoont rat ions of facal calttorma, J. gall Pecal SEreptococcl , Entacoeoeel, 

P- aanjqiiKiaa. and 8tCl.<iot»ctornM sp. rocoueicd firini unXcaty sai/arf* >s wbU u niqn-pnarity am 
non-pnority mami sewaqe during two dry weatJiar aurVBys In IMB. 


Dry WaaUar 
June. 19a 8 

Site r«cal 

Smltflty Sewaqe 

D 9.77 X ID* 

t 1.00 X ID* 

r j.ji X 10* 

HiqR Priority 
Stars Stwaqa 

5. as 


son -priority 
5 ton Sawa^e 

X 10* 

X 10^ 
X 10^ 

«.fl X ID' 

•.SI X le^ 

3.11 X m* 

s.»* X 10^ 

J. '2 X la' 

).M X 10' 

S.57 X 10^ 


1.74 X 10' 

i.>a X 10^ 

I. SI X iB* 

«.ta X 10^ 

S-3S X 10^ 

7,4J X 10* 

I-S4 X 10^ 

X 10* 


l.*5 X Id' 
J.7S X 10* 
t.4< X 10' 

1.31 X 10^ 

4.33 X 10' 

5. 77 X 10* 

1.01 X Lfl' 

••ruQinasa X 10* 

7.0a X 10^ 
1.30 X to* 


3.75 I 10* 

3.11 X lO' 


X 10' 

1,78 X 10- 


l.S» I 10* 

Dry Haacliar 
AUquiC . 19B8 

Santcary Sauaita 


t 1.0s 

F 1.43 

Hiqh Priority 
Stora Sauaq* 

1.31 X 10' 

1.00 X lo-" 

S.64 X 10* 

5.41 X 10* 

Hon -priority 
Stora :IauaqB 

X 10* 

1.3S X 10^ 
t.lO X ID* 
9.S8 X 10' 




X 10'> 



X 10* 
X 10' 
X -.d1 

4.46 X 10' 

J. 30 X :o^ 

T.S9 X iO* 

5,J7 X la* 

3. 03 X lo' 
3.:i X 13' 



4.07 X 10" 

2.S7 X 10^ 

i.ti X 10* 

s.js X 10* 

2.63 X 10* 

X 10< 

1, 14 X 10^ 

X 10^ 
X ID* 

:.8S X 10^ 
3. so X 10* 

in 1983. Kator and Rhodes (1988) also used these organisms to 
diCferentiate human from animal sources of pollution in shellfish 
growing waters. The species of Bifidoba cterium that are 
reportedly human specific and sorbitol fermenting are S^ 
adolescentJB and B. breve . In this study we were able to isolate 
B ■ brave and B. bifidum from human fecal material ; however, we 
also recovered B. adolescentis from dog fecal samples and 

B. breve . B. minimum and B^ t^eoigPhil^m from chicken feces . 

Twenty-one isolates that were thought to be Bifidobacterium on 
the basis of their morphology were recovered from eight different 
sewage samples. Of the 21 isolates, only two could be identified 
by biochemical testing. The two were found in the non-priority 
storm sewer at site Z and were classified as mannose + and 
raannose - strains of B. therm pphilum. 

Based upon our prior use of restriction enzyme analysis to 

distinguish between different strains of Klebsiella pneumoniae 

(Seyfried et al. . 1989), it was felt that genotyping might 
assist in determining the source of the p jf idobacterium strains 
under investigation. Four different enzymes were used to digest 
whole cell DNA from B. adolescentis and two pjf idob?gt.tria35 sp. 
isolated form chicken feces. The restriction (REA) patterns 
demonstrated by the total cellular DNA restriction enzyme 
analysis are shown in Fig. 2. As might be expected, the chicken 
fecal isolates appeared to have very similar patterns. Bam HI 
seems to be the enzyme of choice for the digestion of 
Bifidobacteriuia , and it will be used in all future genotyping 


Figure 2. Agarose (0.7t) gel electrophoresis of total cellular 
DMA from a. adolescent i 51 and BifidobacteFi \M fecal isolates. 

Lanes 1 to 3 contain whole cell DNA from B. adolescentj p digested 
with Sma I, Bam HI and Cfo, respectively. 

Lanes 4 to 6 contain whole cell DNA from a chicken fecal isolate 
of Bifidobacterium digested with Sma I, Bam HI, and 
Cfo, respectively. 

Lanes 7 to 9 contain whole cell DHA from a Bifidobacteriu m 
isolated from chicken feces digested with Sma I, Bam 
HI, and Cfo, respectively. 



Serotypinq of the P. aeruginosa isolates showed that 
serotype 6 predominated in the priority and non-priority storm 
sewage, sanitary sewage and storm water runoff. Serotypes l, li, 
4, 3 and 2, although not as prevalent as 6, were also common in 
all categories of samples. 

Seventy-eight strains of P. aeruqinpg a from the sanitary 
sewer and 113 strains recovered from the storm sewer and storm 
water runoff were also genotyped. Forty-six different REA 
patterns were noted among the 191 isolates. As may be seen from 
Table 3, there was an interesting distribution of patterns among 
the sample groups. For example, REA patterns 1', 6, 6' and 13' 
were found among isolates from the three sampling sites in the 
sanitary sewer. The fact that these same patterns or genotypes 
were also prevalent in the priority storm sewer samples suggests 
that they may be typical of human fecal isolates. A comparison of 
the corresponding serotypes for each genotype showed that the 
serotypes tended to be widely distributed. For example, the 1' 
genotype was found in serotypes 1, 6 and 10; REA pattern 6 was 
distributed eunong serotypes l , 3 , 4 , 6 and 11; 6 ' occurred in 
serotypes 1, 6, 9, 10 and 11; and the 13' genotype was found in 
serotypes 6 and 10. Because it seemed unusual to find such a 
high number of genotype 6 isolates in the non-priority storm and 
runoff samples, the source of these organisms was examined. It 
was found that the bacteria were all isolated during the wet 
weather sampling in July, 1987 from sites X and P. The P site was 


TABLE 3. Distribution of the prominent Pseudomonas aeruginosa REA patterns 
among sanitary sewer, priority and nonpriority stor» sewer and 
stora water runoff samples. 

Konpriority Storm Storm Water 
Sewer Sites Runoff Sites 

X,Z P,Q,G,R 

29 35 

(0.0) 2 (5.7) 

5 117.2) ID (28.6) 

(0.0) 1 (2.80) 

(0.0) (0.0) 

4 (13.8)* 6 (17.1) 

2 (6.9) 3 (8.6) 

^ Total number of REA patterns or samples in each category. 

• Found at all sites in the sample category (e.g. sanitary sewer, sites D, E and F) 





Sanitary Sewer 
sites D,E,F 

Priority Storm 
Sewer Sites 




























an additional street runoff sample taken at the beginning of the 
rainfall event. These isolates, belonging to REA pattern 6, were 
evenly divided between serotypes 1 and 6. 

Additional information not provided in Table 3 is that 
genotypes 1, 2, 4, 7, 9, 11', 12 and 14 were all isolated from 
sanitary and priority storm sewers and not from non-priority 
storm sewers or storm water runoff. 

In comparison, P. aeruqlnpg a isolates with genotypes 18 and 
20 were found solely in the non-priority sewer and storm runoff 
samples. The REA pattern 18 organisms were distributed between 
serotypes 3 and 6, whereas pattern 20 was found in serotypes 1 
and 3. Serotype 3, REA pattern 18 P. aeruginosa isolates were 
found in the storm water runoff samples at site Q, and in the 
sewer that collected this same storm water at site Z. Similarly, 
REA pattern 20 isolates were recovered from water runoff at site 
R and in the receiving storm sewer at site X. The fact that the 
genotype 20 organisms belonged to both serotypes l and 3 suggests 
that gen oty ping is probably a more concise method of 
fingerprinting P. aeruginosa than serotyping. From the results, 
genotyping appears to be a promising method of tracing p, 
aerutjinosa from human and animal sources. 

Compared with P. aeruginosa , genotyping of the fecal 
streptococci did not produce any concise results. One hundred and 
ninety-two streptococcal isolates, 60 of which were from the 
sanitary sewer, were genotyped. A total of 64 different REA 
patterns were identified among the isolates. 


streptococcus f aecalis subsp. faecal is was the only species 
that had the predominant genotypes 5 and 8 occurring in both the 
sanitary sewer and the priority storm sewer isolates. Mundt 
(1973) has suggested that s, faecal . js isolates from human feces 
will produce an acid curd in litmus milk. However, no 
relationship between the 5 and 8 pattern isolates and those that 
produced an acid curd was observed. Although the S. faecitfT CT 
isolates were distributed among 31 different REA patterns, 
genotype 9 was found in all three sites of the sanitary sewer and 
genotype 5 was recovered from all sites in the priority storm 

sewer . Sj faeciua subsp . casseliflavus had 2 1 different REA 

patterns that were widely distributed among the isolates from 
different sources. In general, no relationship between the source 
of isolation and the genotype could be found among the 192 
streptococcal isolates studied. 

More insight into the origin of fecal wastes in storm sewer 
1 ines was provided by the speciation of fecal streptococci , The 
results showed that S. f^eciv i p tended to be equally represented 
in all sample categories. On the other hand, S. faecalis subsp. 
faecaj-jg (Fig. 3) was found more frequently in sanitary and 
priority storm sewers than in surface runoff and non-priority 

sewers. In contrast , g^ faecium subsp . casseliflavus ( Fig . 4 ) 

predominated in non-priority storm sewer water and was notably 
evident in storm water runoff. The organism was virtually 
nonexistent in sanitary sewage and levels in priority storm 
sewers were small. These results concur with our previous data 





A B C Y 

P G R Q 

Fig 3 Percentage distribution of Streptococcus faecalis subsp. faecal is among the sanitary sewage sttss 
(D.E.F). the priority stonn sewer (A.B.C.V), the non-priority storm sewer (X.Z) and the storm 
water runoff (P,G,R,Q) locations. 



Z 20 



' * 

D E F 

A B C Y 

X Z 

P G R Q 

Fig. 4. Percentage distribution of Streptococcus faeci ^ imi subsp. casseliflavus among the sanitary sewage 
sites (0,E,F), the priority storm sewer (A.B.'c'.Y), the non-priority storm sewer (X.Z), and the 
storm water runoff {P.G.R.Q) locations. 

(Seyfried, Harris, Young, 1986 unpublished) which showed that S. 
f aecium subsp. casseH f jiflvijg could be isolated from animal feces 
but not from human fecal specimens. 


The conclusions that may be drawn from this segment of the 
study are as follows. 

1. The levels of fecal coliforras, Escherichia caii, L 

aeruginosa and Bifidobacterium sp. suggest that there is an 
impact near site A in the storm sewer line that may be due 
to human fecal pollution. 

2. While B4f i -Jp feacterium breve and b. adolescentis are found in 
human feces, they cannot be used to differentiate human from 
animal sources of pollution because they can be isolated 
from animals such as dogs and chickens. 

3. Genotyping of Bifidobacterium isolates may provide a more 
precise method of source differentiation. 

4. P. aeruginosa genotyping may be of value in tracing sources 
of pollution - Se retyping , however, does not provide 
specific results. 

5. Genotyping of fecal streptococci is not recommended as a 
method of source determination since the wide variety of 
patterns produced yield inconclusive results. 

6. Speciating fecal streptococci is a useful means of 
characterizing sewer or storm water content. For example, g^ 
faecalis subsp. faecal is is found predominantly in sanitary 


and priority storm sewers whereas S. faecium subsp. 
casseliflavus is characteristically present in non-priority 
storm sewers and storm water runoff. To date, S, t a^ffi wni 
subsp. casseliflavus has not been isolated from human feces. 


Bradbury , W. C , , A , D . Pearson , H. A . Marko , R . v. Congi and 
J.L. Penner. 1984 . Investigation of a Campylgbacter ieluni 
outbreak by serotyping and chromosomal restriction 
endonuclease analysis. J. Clin. Microbiol. 19: 342-346. 

Bradbury, W.C., R.E.G. Murray, C. Mancini andV.L. Morris. 1985. 
Bacterial chromosomal restriction endonuclease analysis of 
the homology of Bacteroides species - J. Clin. Microbiol. 21; 

Dufour, A. P. and V.J, Cabelli. 1975. A membrane filter procedure 
for enumerating the component genera of the coliform group 
in sea water. Appl, Microbiol. 29: 826-833. 

Dufour, A. P. 1980. "A 24 -hour membrane filter procedure for 
enumerating enterococcl". Presented at American Society for 
Microbiology Annual Meeting. Miami Beach, Florida. Hay, 

Dufour, A. P., E.R. Strickland and V.J. Cabelli. 1981. Membrane 

filter method for enumerating Escherichia coli . Appl. 
Environ. Microbiol. 41: 1152-1158. 

Geldreich, E.E. and B.A. Kenner. 1969. Concepts of fecal 
streptococci in stream pollution. J. WPCF. 40: R336-R352. 

Kator, H. and M. Rhodes. 1988. Evaluation of alternate microbial 
indicators of fecal pollution in a non-point source impacted 
shellfish growing area. Abstracts, First Biennial Water 
Quality Symposium: Microbiological Aspects. August 29- 
September 2, 1988, Banff, Alberta, p. 6. 

Hoadley, A.W. 1977 . "Potential health hazards associated with 
Pseudomonqs q^ruginosa in water". in Bacterial 

Indicators/Health Hazards Associated with Hater . Hoadley, 

A.W. and B.J. Dutka (eds) . ASTM Special Pub. 635, 
Philadelphia pp. 80-114. 


Kusama, H, 1978. Serological classification of Pseudomonas 
aeruginosa by slide agglutination test. J. Clin. Microbiol. 
6(7) ; lBl-188. 

Mara, D, D. and J.I. Oragui. 1983. Sorbitol fermenting 
bifidobacteria as specific indicators of human fecal 
pollution. J. Appl. Bact. 55: 349-357. 

Mundt, J.O. 1973. Litmus milk reactions as a distinguishing 
feature between streptococcus Caecalis of human and non- 
human origins. J. Milk Food Technol, 36(7): 354-367. 

Sey fried, P.L. , E.M. Harris, I-J. Huh, R. Harmandayan and 
E. Hani. 1987. Characterization of the fecal indicator 
bacterial flora of sanitary sewage with application to 
identifying the presence of sanitary waste in storm sewers. 
Proceedings Technology Transfer Conference, Part D, 
Analytical Methods, November 30 - December 1, 1987, Toronto, 

Sayfried, P.L., R.M. Desjardins, A.E. Alarcon, N. Kulendran, 
H. Sidarous, E. Harris, w.c. Bradbury and M. Young. 1989. 
Antibiotic and toxicant susceptibility profiles of clinical 
and environmental Klebsiella pneumoniae isolates. Toxicity 
Assessment, ^ (in press) . 

Slanetz, L.W. and CM. Bartley . 1977 . Numbers of enterococci in 

water, sewage and feces determined by the membrane filter 

technique with an improved medium. J. Bacteriol. 74: 591- 

S^iandard Method;? for tf^e E;?;aiT|j ,ptit;j ,Qp of Water and Waste-water 
16th edition. 1985. WPCF. , APHA, and AWWA (Eds.) American 
Public Health Assoc, Washington, D.C. pp. 978-979. 




J. Hog*'- Pitblado and D*1b a. Dcmpuy 

Department o* Gtograony 

L«orer>tian universitv 
SuaOury, Onta'-iO P3E 2Ct» 

Qigitkl cl*t« from the Mul t l«p«c tral Scanner (1^51 o1 tl-t* u*rtctsat 
evits o1 sat«lli.t«s r>Ave bvvn vniployed since the launch of Lana«at-1 
In 1972 tor a xide rang* of la^e Mater auality assv^xnB-'t progfamt 
(B'oalts, lf75; Fisher eC «1 . . l")??; Scarpace *t al., 1979; nU»«ana 
et al . , l''B3i Vera in, 1985) . Thos* ac ti vi t ie» have only aeen 

partial ly succesvf ul Que to the relatively coarse Spatial, ftpec tral , 
anB radiometric resolution Q* flSS data IMlddlPton and Wunda., Jr., 
19B0; Witnq and Whitehurftt. 1981! Hilton, i^Blj PltUiado, 196*! 
Lmdell, 19B&1. But «ith the launch of Uand«ats 4 and 5 in 1982 »na 

IfB* , respect I vel v , ftatet) I te water qual ity asse&tmen ti have been 
enhanced because of the foil Owing significant Oesign improveinen tfi o* 
the Themat ic Mapper scanner (after Lathrop, Jr. and Uil leB«nd , 1986 ( i 

o 30-n. versus BO-™ ground resolution in the visible 

and reflected infrareo Pdndsi 
□ seven bands of sensing versus four banOs, notably 

with a new band in the blue wavelength region aod 

a thermal infrared band: 
O B-bi t ver«u« t-bi t radiometric resolution , with Che 

consequent 2 54- level recording of data compared to 

A4- level . 

Iti the late*.t issue of Pnotoorannnetr ic grFqineer mo and Kemat» 

Sensing , another satel 1 i te monitoring tool for water qualit* 


«*4psament» 15 r*Dort»o an - tii» Frenrr' SPOT o*5»llilP, I fluicr^rd C^ 2l 
February 1986. Mffrry et <1. (I'JHB) (JiBCuo» the usa o1 the ICi-o. 
mul t ispsc tr*| rnrjtlo of SPQT mR^ data 'or ntapplnq raldtivc ranqe* o- 
vuspBndad »»d*m»nt concent rat lor>5 i" La'tp e--i«. U^'ortunjltly, 

daspi t» t hr increase in Bpjt lal re^olut 1 ori , SPOT' dat.a <i>>: t 1 nave? 
1 iffli t*d appeal to nttor martaqar^ ariiS scientists OecausB a1 thv lac^ at 
* bi»na jn the blue par t ion of tne apectrum. 

3Cudie« tn Nartheactrrn Oritarto using •jitvlli.te inagffry ^or Matv 
quality a^sesani^nts l>ave ^ocu«^#d O" the u«e o^ Lanoiat oigitdl data, 
wl tti Tio* t success cominq from ttip use a1 TH as □DposeO to MSS scannpr' 
data. In feasibility studies dealcjried to discriminate between c loar 
acidtc lakes -from non-acidic lakes, cla»si f icat lona in tie order or 
90V, correct have tj^mn acnicved (P^tbladtj et *l., l*?B7i Pitbl«ClO, l''87a 
i<?871j and 198BI . 

Expanding oe/ond the feasibility atudy of I9B6-B?, tfie author*, 
are no™ er^gaged m a thr-ee-year rt^tart^i' pi-ojecl funded t>v tfie Ontario 
Ministry of ttn« Environment IRfiC Project No. 35flQ I . The general aim 
jB to c fiarac ter I le and map the lakes in thrpe selected arra^ 01 
Northern Qntarin u»ing Landsat Til data. Thi« p»amr d>>*tc r 1 be« iome of 
tne work uf>Oer taken in th» firat eight rnonths of that study. The 
Objective today is to describe the nature of the TM data that have 
b*<rn acquired far Northeastern Ontario and identify a limited number 
of aaaociations betweeri Chose dala and a selected Mater Quality 
parameter, riorn specifically, use is made of the techniques a' 


principal components analv5,B5 ( PC« i and discriminant jnalyees c- TM 
dat* {iron. LanOaat-5> in association with measured values of ai5SO.,ea 
organic carbon i DOC 1 . 

Stuflv ftrea and Lanflaal; ImaciiTry 

Spectral responses acquired Dy the Lana»at-5 Thffmatic Macaer 
scanner ( TM ) are being gathereO (or all water bodies larger than one 
hectare (it they are detectable by the satellite) in the art-^ of 
Northeastern Ontario that extends from the North Channel of Lake H^'on 
to Highway 11 { south-nor th J and fro-n Wawa to Temaqami Iwest-easl), an 
area of appra« imatel y 120,000, The region is roughly outlndd 

by the location of 635 of the stud/ lakes in Pigure 1. Giver -.he 
scale of the map and th» small size of many of the lakes, some of tt-e 
dots there may represent the location ot up to two of the It^ee 
sampled for analyses. 

For the purposes of discussion here, the mean vaUies of the TM 
response* for all of tHe seven TM channels for each of bZZ lakes r-.ave 
been employed. The Til channels include: the visible bands TMl i-50- 
520 nm), TM2 (520-600 nm ) . TM3 (630-690 nm 1 ; the near- to mid-infrared 
bands TM4 (7AO-9001, IMS (1550-17501, TM7 (20G0-2350 nm 1 i ar.d a 
thermal channel TM6 (101O0-125O0 nm ) . Because a relatively s 1 ol« and 
sire- 1 1 mi ted microcomputer ( the IMAVISION system of PCI Inc. of 
Richmond Mill, Ontario! was employed to acquire these spe; r 'al 
statistics the authors designed a -lamoljnq program and computer 


proceou'^s that wou 1 remavw atmospfier ic na;e (Richaras, 19861, 
eliminatH rion-Mater pixels, per(o'-ni a principal comoonefit* analysis on 
tliB "water- pincls, outlinv the water bodies with a pol vgon-border , 
and then cpnipute and record the descriptive statistic* for each of the 
sampled water bodies. fls the study area is very large, t-enty-ane T(-; 
quadrdits Mere used from the Landsat-S images listed in Table 1. The 
routine described above was applied to a quadrant from one to five 
times loepending ori the location and distribution o* candidate lake*) 
«ith aact-i application requiring appron imatel y two to three hout-s of 
automated and interactive processing time. 

Figure 1. The study area is outlined by the location of 

the lakes in this sketch map of the northern 
portion o* the Province o* Ontario. Note the 
t«ck of study lakes in the Tioiniins »raa Hr\are 
no cloud-free suiMoer imagery mas available. 



Listing Q< the LaridEat-5 Ihemat it Mapper Images 
Used in the Analysis a1 LaUv Spactr-^l Responses 
yn Nor t>ieaatef-n Ontaric3, No single year coulC 
be employed betause af varying cloud cover but 
the images selectpc were restricted to July ano 
August - to match the lake water quality sampling 
dates . 

Date □■* Image 

Scene l.D. 
































154 40fa 





Ful 1 


5051 a 




























Water Parameter Data 

Water parameter data for the sanipled lakes were compiled fron- a 
number of sources but the oDjective was to obtain these data from 
lakes that were sampled within a Julv~'^uguat window as close as 
possible to a Landsat overpass. 

This abjective was met for a large nuiTpber of lakes in the Sudbury 
and Alqoma Ar-^^^ because of the field work undertaken by the senjnr 
author in 1986 (Pi tblado, i9a7a ) . For those lakes, which make up 

c lose to 50V. of the study lakes, the difference in timing between 
Landsat data acauisition ana field data acgu i si I ion ranged from a few 
minutes to one week. However, the remaining data acquisitions 



d«p»nd«tl on AccVkstng th« d»t«bdees a* Cne ni'^istries of Envlronman t 
«r>cf N*tur«l RsBourcVG ( •»> Acknow ledgemsntB t ana fnatcriinQ with cloua- 
1 raw Landsat imagery. Consequent! v the timing differences for •onie of 
the remaining study lakes couliJ Oe ds iarge a& one full year. 

In effect, four lake dacabAse« (noting that the four do have 
•Ignlficant overlap due to the sharing of data between liinl«trie»; 
also, for analytical nethodG aes MQE , 19911 were employed: 

Pitblado/rnce, i'?B7ai the MOE fl.P, I .Q.S. datasota described by 

Pitblado end Keller (19S4J and Keller and PitDlado (I'^Bbl; the Inland 
Lake Oetabase fro* MOE Dorsetj and the ftquatic MaOitat Inventory of 
the Ontario Ministry of Natural Resources. The water parameters (and 
SOMe of their descriptive slatistictl that were acquired from these 
databases arg listed m Table 2. That table hxghliqhts sone of the 

unevenne^s in parameter acquisition and , in the opinion of the 
authors, ts a further argument for the use of remote sensing to gather 
data for selected water parameters in regions nhere lai^es are 
numerous, remote in location, and scattered over an enormoos 

geographical are*. 


T*b!» 2. 

0»%CriBtive Stativtics for lUe Lalto WatBr 
Parameters Compiteo for this Studv- 

Hvan ( n } 
























22. OOO 





























400 , 000 











1 24 . 000 


















30 . 000 










7. 100 








1 .70O 

4 24 






34 7 





















































DOC Iraq /LI 

Lake area ( ha ) 

Lake depth (m) 

Chlorophyll a (ug/LI 

SeccH) d»pth l«) 

Apparent colour (haztn _nits) 


TIP ftikalinity (nig/L) 

TFE Alkalinity (ftiq/L) 

CohduC t J vily (yfflhoi'CT") 

TKN (ug/L) 

NH3 (ug/L I 

N02 ipg/Ll 

ND3 (uq/LI 

Total P (ng/Ll 

CA img/L) 

MB [mg/D 

NA Img/Ll 

K {mg/D 

SC4 [mg/LI 

Si03 («g/Ll 

CL (mq/L) 

AL I ug/L) 

MN (yg/L) 

FE fug.'UJ 

ZN lM9'Ll 

CU (wg/Ll 

Nl (ug/L I 

PB (jig/Ll 



To f ina ttie under lying aimensiont o* rsmately sefiBeQ Oata, a 

comnionlv used technique is that o1 pr i nc i pa i compone- r« analysiB 

(PCfl). A» describsa by nichams fl'Jat) anU characteniBO =■ Pung end 

LvDrvw {19B71, the PCA transformation involve* thr»8 Btapsi 

o derivation o* th« vari«nce-c ova r j ante ma t rn- 

O computation of •ia»nv»ctor» 

o linear transformation of the dataset 

it has long b*«n recognireo that mu 1 tispectr* I data < rem remote 

aifhsinq scanners o* th» Landfiat series of Batellitea (as well as from 

Other scanners with similar band selections) exhibit hiqn interbanO 

corr»l*tions and tMcrefor^ may involve a consiOeratjl* degree ct 

r»dund«ncy. Thus, the uncorrelated tin*ar|y transf nrmeo comoonents 

are derived from the original dataset in a manner such that the first 

princip*! component accounts for the maximum possible p-'aportion of 

the variance a* that dalasot. Tne remaining comporents, in a 

cl««c»ndlng ordered ■•quence, account for the maximum proportion of the 

unexplaineo residual variance. With Landsat MSS and TM data it ia not 

uncommon to t ma HO* to 90» percent of the enplained variance in the 

reflectance dataset to be found in the first or first plus second 

principal components . 

Virtually all applications of PCA to -emotely sensed data have 

been employed to assess terrestrial I vegetation, geology, etc . I 

target*. An enCBllent review of some of the more significant uses of 

PCA far much purposes, as hsI I as a discussion of alternative 


AcproacHts within ths t«chniQue it«»lf. i% provlavd &v ^unq «nd L*Dr*M 
II9B7), Our- papwr , perhaps the first of it» kind, focijSe* on only 
dquatit targets, the lakes of No' t heas'.ern Ontario. 'ti« orinciEJal 
data&et tonsisted o* the mean values ot tr-e seven TH bands *or each of 
the 1a^«s in the study area. This wntire a«ta«et uas «uBjected to PCA 
a& a whole Or was subdivideQ prior to PCA u«ing d number o' i;ategories 
based on selected values of DDC . All seven eigenvalues Mer^ computed 
for pacn subset af batd but only tnp statistics tar the first five 
principal components are listed for tne illustrations Delow- 

Tne eigenstruc ture of th<- TK data basvd on alt mean pmel values 
for the &33 study lakes is provided in Table 3. Prom that table it 

can be seen that the visible baridc TK3 and TM2 are heavily luaded an 
the f irst component <FCA-1 ) *ncl the visible Iblue) band TMl i» 
inod«r«t*ly loaSed on that same component. The mio- i^ fr area bands, TM7 
and TM5, are heavilv loaded on PCO-2, and the near- ir t rarea band, TM4 
IS loaded liighly on PCA-3. Tli««* tnree eauiponents together account 

for 8B . 2V, of the overall variance of the lakes reflectance dataset. 
They provide evidence of the expected cDntr*»t between tne spectral 
responses ol watw tioQies. This is attriQutea to 5 ne 'act that clear 
water bodies, (or tne most part, absorb infrared radiation but ara 
highly reflective in the visible portion of the spectrum. 

The visi bl e*then-inf rared seouence o* componei t» are in direct 
contrast to many published -orks on the applications of PCA in remote 
sensing. As illust'ateO tiy the work of Fung and LeDrew (1997), the 



no'-fial eiippct*tio" would be ''or arx infrared- then-visi ble saries cf 
componsntB b»catj»e audi narie has focmssB on tprrp»trial targets. 
There the great variety □< vegptation types rcaulti in a PCfl-1 tl>at 
ttiBcn mioate* beti-«en th« fiign infrar»d i-oflecting targvte o* foreat 
*na crop land to tn« |om inf r*r«d r»f i»c ting »urt*ceB of plowed 
fifflds, cutowei-s, or lands in rura 1 -to-urban corve'ision. 

PCA-S, with tr« blue waveband ICIl laaOinQ heavily on it dr.o 
cDnf Ibut ing t ee« than i our percent to the ewplaincd variance, SBems 
sdRiewhat paradoxical. Me have argued, as indeed many have argu»d , 

that the addition of Tni to the Thematic Mapowr la one o* the 
&trenqth« of that scanner for water quality analyses compared to the 
Dana selac t xons aval labia for the Land»«t MSS or the SPOT hRV 
instruments. Why ien t TMl a iBajor contributor to PC1-1'' It would 
apocar that it acts as a controlling variable, in the sense uf a 
partial correlation, when all Mater bodies ara being examined. Later 
in this paper » t wii I bff spen that 11-11 plays a much more significant 
rolo when contrasting water bodies Dt differing DOC. 

Of great interest ta the contributian of the thermal band ( TM6) 
to the fourth most important principal Component (PCA-41. UJhile only 

five percent of the total variarice is accounted 'or in thj» coiTiponent. 
ll does support earlier observations (Pitblado I'JS/a and l-JBTb) that 
clear lakes (i.e. highly reflective in the visible! tend to be cooler. 
It IB a great pity that the TM6 spatial resolution is bo coarse 


(I20-ni). In m*rv ir(»tanc(p% in Nor th«**tBrri Qntai-to ou'- «tiidy Ichvv 
are so cni« 1 t that f^e TIi6 r»5pDri*es ar» too often coming from I»kB- 
lake ahorp niiKel*. It is to be hopwO that future ^latpllite Bca^r.prs 
Mill fu^iaic *iip*i-iO'' tncn-al senginq capaCii I 1 1 ie"» mal <~i 1 1 enanle u^ 
to FHore easily datvc t the mutramely iTiportant differences of Mithin- 
and between- l«k« temper* tur» dif f erenc**. 

Table 3. E igenstruc ture of ail Mater tiodies tr> tl^e study area. 













. 1069B 




. 15921 








. 19464 











. 10662 

. 119BB 




-.094 72 






. 1B5B3 

. 17121 






. v«<?e7 




'/. varia 



13. o 





'/. ti ar lance 

Ad. 5 






In order to display the contrasting PCt- '~esu1ta trie .aVes uert 
Subaiviaot) into cri'-ee subsets Dasec) on arbttranlv selected categories 
of DDC. The 1 ir%t subset, of 66 latiet , cofSiEttfd o* lavee nith a DD' 

concentration of 2.0 ibq/l. or teas. The actual range of DOC for thi? 
group of lakes >iaa from 0.1 to 2.0 with « mean of 1.04 n>g/L and a 0.66 
etandarg deviation . 

The iBcono subset had a mean of 3.06 mg/L and a standarO 
dwlation of 0.52 inq/L. Tnja group contained B8 lakes -iith DOC 

ranging from 2.; to 4.0 mg/L. The final set of 05 lakes ranged in DOr at ion from fl.l to 6.? mg/L . Here, with slightly greater 

Ml thin-group variation as espresied tjy a staiCarO deviation of 0.71, 
ttiB average concentration was 5.13 mg/L . Each of tfieoe subsets o* 

lakes was subjected to PCQ using the same routine as was emciloyed fa'' 
the entire dat aset . The numerical results of those analy^** ^' ^ 

tabulated m Tables 4 to 6. 

□n« would anttcipatB that a group P' lab^a with little or no I -~ = 
2,0 mg/L) dibsol vttd organic carbon would oe highl v ref >ec tive, 
espetiallv i" t"e visible band* of the Thematic Mapper. This is !■ 

(act the case a« illustrated in the »unimary Table 7, noting that the 
highest spectral response occurs in TMl , The eigenstructuro of this 

group (Tflole fli of laties reflects that fact wit-i TMl explaining littl'- 
of the variance of that dataset . There the visinle bands TMl (blue) 
*nd TH2 (green ) load most heavi ly only on the third and fourth 
components, respectively. Ii"' Northeastern Ontario, to enplam ttie 
variance i" these highly reflective lakes one must look to subtle 
differences m the r,e*r - and miO-infrarea bands, partlCularlv- '*s 


eig«nfttfuctijr« of r«tie» in tho study *rea v,jth bt 
!e«ft than or equal to 2.0 mg/L. 













. 09 1 69 









,3<»4 71 

. 2B3&2 












-. :523e 

-. 14342 







. 4B070 

.194 54 



. 16276 


va lueft 




. 18416 


V. uarianc* 














Tablp 5. 

Eigenctructur* of lakes in the study area with DOC 
'i'-'^t^r than 2.0 Dut I r«s than o"- pQual to 4,0 mc'L. 










.3007 5 

. ■^0989 







. J':;405 





- . 26850 








.395 IB 


. 5024H 

.3 7895 



. 3 1 550 


. 70608 












. 1671B 

. 12757 

X varianc* 






Cum . V. 

var lanco 







Table 6, Eiguriwtruc turs a* lakes in the itudV arwa with DOC 

great«r t.nan fl.O but l>as tfiart or equal tc b.^ mq -. 



















. 252G7 



. )'li44 

.4 2693 






-.11 3S9 





, 15J111 

. 82685 





















'/. varLance 






cum . 

V. var i«nce 

77. B 

99. J. 




Table 7, Mean values i Haie-co>-r»i: tea digital numbers) for each 

of the «»v«n TM banda for eacH of t^ie three DOC qroupa. 


Them«t kc Mapp>r Bands 
TM2 TM3 Tn4 TM5 

11 .05 5.13 4. 74 9.04 6.B4 121.58 2.98 

7.70 4.35 4.fae a. 20 6.82 121.81 2.95 

6.66 3.82 4,45 7.50 6. 17 121 .84 2.92 

Table B. ClaEBif ication result* L.Bing d iBcr iminant a^alvsiB- This 
analysis employed the thre» DOC-defined groups and the 
»ej»n TM bands. Included in fits table are the predicted 
group membersnips of the 434 lakes «hic« -here not 
included i<i th« predtf inad groups. 

DOC No. of 
Group CASES 

Predicted Group Me 
1 2 


1 (Low DOC) 66 

2 88 


78 . 9V. 


6. ax 

I'S. 7V. 



1 . 57. 


39 . BV. 

3 tHigh DOCl 43 

4 . 47. 

1 7 . BV. 


7 7 . 67, 






muc" «B BV'. of the total variance is •>>pl«in«a by tl-io«« TM BandB that 
centre around the near- in t ra'-ed i Ti-ia and TM51 , inc luding the vi%iOle 
band TM3 I red ) which tend^ lo overlap "ifi thp near- i n frared . A minor 
con t*- i butian ta this component i« pro.ided Oy the mid- in f rared band 
( TM7 ) . This latter band is really a* little significance a* suggested 
by the mean v«lu«S of TM7 in the thr»e DOC groups Of lake« (Table 7) 
which differ only by a tnaxiinum of .Oii haie-correc ted digital numb>r«. 

Slightly more than seven percent of the variance within low DOC 
lakes IS e'plained by the PCA-2 where the thermal band ITn6) is loaded 
heavily. In tact this band ahOMS up on the second component of all 
three Iflkp groups defined by the DDC categories, enplaining from 5 . "f Z 
(Table 51 to 1 1 , 2"/. (Table 61 of th» total variance «ithin tie 
respec tive lake groupings. Frankly, this i» somewhat surprising. 

While we argue the value of ttiis therina] band in looking at water 
bodies in Northeastern Ontario, it is clesr from Tacle T that thn 
digital nui^bers of TMii vary little f'-om one group of lakes to another. 
This spectral ri?sponse and its apparent ability to dtscr iminate 
bwtween various lake types wi 1 L require further investigation . 

Turning to the other entreme of the DOC groups with measured 
concentrations greater than 4.0 but le*5 than *.5 mg/L, we see that 
the eig#n«truc turc (Table 61 is the reverse of that for the low DQC 
group. There the visible bands ( TMl , TrtZ . IMS) are loaded highly on 

PCA-1 and it is the near- to m id- infrared bands ( Tfia , TM5. TM7) that 
occupy positions of heavy loadings on the third and subsegurnt 
components. find again the thermal band is the most significant 


v«rigible to 1 o*ct on PCft-2. 

We attribute this reversal of t^^e loadings Co the tact th»t the?.! 
arp tfie mast coloured of the lakes in the entire Qataoet. They woul" 
b* enoBct^a to rsflact mor-v in t^ie infrared in comcdriKon to the 
vieiblp bandft. As euggasted by the <igurff« providwd in Tabl* B, these 
^^ompar 1 sons must be made :T>o5Cly in relative and not absolute term^ , 
Thus, in the enaminatior of thp eigenstruc ture ITable ?i of the hjgr 
OOC lakBs MB sec that if th«s« lakea ar« to b« differentiated xithi- 
that group one Hould have to BNaniln* th» visible bonds nhich CKplclf 
77. S'/, of the variance. 

Our middle qroup, with DOC between 2.0 and 4.0 mg/L, ia ma^ ■ 
similar to the higher DOC lakes. Again (Tabl« M. the sequence of 

cofioonen ta and the respective loadings of the Tm bands arei PCA-1 , 

visible bands) PCA-2, t tier ma 1 band; and the infrared bands loaded on 
the third and subsequent components. The visible band that occur --. 

very close to the intrarefl, TM3 , plays an insignificant role. 

We htv9 conduc ted an eKperiment to sea whether classification 
using discriminant analysis I SPSS I nc . , l^BH ) would maintain tnp. 
•tructure of these DOC groups. Summary results ar^ provided in Table 
a. That taRle banically preserves the interpretations suggested fo-- 
the eigenstruc tures of the t.r^rea DOC lake groupings. Cloce to eighty 
."pitrten t of the lak»o in the first and tnird DOC groups ar» correc 1 1 , 
classified, but the lakes of the second group are spread primaril, 
between the middle (setond group) and the high IthirO g'-Qup) DOr 
lakes. On the canonical discriminant functions that Mere generated 


for thiB *ialv»is; till and TM6 viere the only banOs Mith niqnif»t«rt 

correlat ior\« on th^ f irst func t ion , account inq iar 9c . 1 ''. 3' f-p 
variance (P ". 0.001). All of the Qther TM bandH were cor-plated, if 

al all. wi '.ti the second discriminant Junction. 

Giver, the fact that our DOC group limits xere defined 
and that i«.B have not employed any numerical traria'ormal ions of the TM 
data *•} 1^ so often the case far water quality analy^KS lE^arcace et 
al., 1979; ^erdln, l9B5i Lathrop. Jr. and L i U esand . l-JBt). we are not 
tli*plea5«d Kith these preliminary results. Indeed, it is iiteresting 
to note that of the A3* lakes identified in Table 8 as UngrcjOWd. Only 
two have DOC measurements that xe can employ for comparisc" Durpose*. 
The»« tHQ lakes have DOC concentrations of 9,7 and 10.5 mq/L, 
respectively. The discriminant classification showed that their 

membership of highest probatJilily would be group 3. Eifact./ as woultj 
b» enpectt-d Por comparison purposes with Table 7, the banz rreans for 

these two lathes are I from TMl tn TM7 , respectively)! -1.62, Z . 10 , 3.2S. 
5.90. 4.48, 122. B3, antl 2.01. 

This summary, interim paper has described some of the basic, 
underlying structure of the Landsat-S Thematic Mapper spectral 

r-e5pQO*.B6 of lalie^ in Northeastern Ontario. The Oesciption and 

discussion has been provided for the entire, current dataset as mbI I 
as subsets based on DOC lake groupings. F^uture wo'k will be 

undertaken for tupset* of the sampled lalies -here the gro.jpinys are 
based on other optically significant water quality para-tieters and 
predictions made for those parameters for all lakes in the study ^raa 
that can be discriminated {"lacated") by the TM-5 sensor. 



This researc h hat bf-en supported by an RAC ( MOE ] gi-ant to J.R. 
Pitblado with appreciation e.ttnOeO to ll>e outstanding as5i3t«ri(r» froT, 

th« Project Liaison O'ticer, W. Keli»r of the Sudbury office a1 the 

Minis-try of the Environment. The authors are a 1 ao grateful for the 

provision o* data I'-om the databases of thw rKnt^try of the 

Environment iDave Gardner, NDE Dorsetl and the Ministry a1 Natural 
HeBoarces ( Donna Wales, MNR Toronto I . 


Brooks, D. J . 197 5. Landsat measures of water c 1 ar i ty . 
Ph ptoqr#mnietr|li: Enoineer j.r>o and Hemotg Slfhsinq , flO: i 269-1272 . 

Fisher, L.T., 5carpace, F.L., and Thomson, R.G. 1979. Multicfate 
Landsat laUe quality moni tor ing program, Photagrammgtr i C 
^nqine^rrinq and Remote SenBing a5!51''-527. 

Funq , T. ant) LeDrew, E. 19B7. Application of principal components 
analysis to chang* detection. Photogrammetrn: Enaineerino and 
Remote Sensinn S3il*.'l9-165B. 

Hilton, J. 19B4. Airborne remote sensing for freshwater and 
•Btuar ine moni tor ing , Ulatgr Re^earqh 18: 1195-1223. 

Keller, W. ano J, Roger Pitblado. l'?8*>. iiater quality changes ir. 

Sudbury area lal«es: A comparison ai synoptic surveys in 197a-76 and 

1991-83. Water, Ai.r , i^nq Soil Pollution 29:2B5-29fe. 

Lathrop, Jr., R.Q. and Lil lesand , T .M. 1986. Uao' of Thematic 
Mapper data to assess water Quality in Green Bay and Central Lake 
Michigan, Photogr ammetr jc EhQinyprinp anff nemote Sgnsinp 

Lillesano, T.M,, Johnson, W.L-. Deue 1 1 , R.L., Lindstrom, O.M., 
and Meisner, D.E, 1983. Use of Landsat data to predict the 
trophic state of Minnesota lakes, Photooraminetr ic Engj hearing 
and Remote Sensing fl9i219-229. 

Lxndvl I . L. T. 19ab. Opera t lonal nater qua 1 i ty fturvei 1 lance in 
Sweden using Landsat MS5 data. Proceedings ■ Tenth Canadian 
Symposium on Remote Sensing, pp. 3B5-394 

Herry, C-J-. Mc^Um, H.L., LaPotin, N., and Adams, J.R. 19BB. 
Use of SPOT HRV data in the Corps of Engineers dredging progi-atn, 
Photogrammet ric Enginqerinq and Remote Sensing 54 : 1295- 129''. 

Middleton, E.M. and Munday , Jr., J.C. 19B0. Landsat - What is 
operational in water resources'" Proceedings. 5i n th Canadian 
Symposium on Remote Sensing . Hal i f a« , pn . «3-52. 


Ontario Ministry o* trie Environment. i'JBl . Oi^~ltng» Q* 
flrijl vt 1C*I WethoflS - Torontoi MOE . 

Pi tbl«da, J ,R. . 1984. Monitoring tne imtiac ts it airborne 

pollutants with digital reinotB B.vns.ingi progrp^^ ana pro^p^cts 
m the Sudbury ara». \.« ki^ r^ r\tl»ri Univernty Beviaw lb 167 -71 . 

Pitblado, J.R. and Keller, M. 1^94. M gnitgri-j o< Northeastern 

Ontario La><g8. .i,y 9 J-Ji,yg?. Data Hcoort. S\jatfjr , : Ontario 
Mini»trv O* the Environment. ( 34B pp. > . 

Pitblada, J.R., T«ni*. F.J., and Woito«ich, w.h. lfQ7. «»*□(» 
aeniing and Che monitoring of lake ac idJ * icatic-. in Northeastern 
Ontario, Canada: Prel iminary abservationa , pp. 3<>fl-'71 , IN B- 
Perry, R.M. Harrison, J -N.B. Be 11 and J.N. Lester (ed». J 
flc ^d Rain; 5c icnt 1< ic and Typnnfc^l Advances . _ and on i Engl and) ! 
Publication* Division, Selper Ltd. ( ProcegJinqs . International 
Acid Hiin Conference, Sheraton Hotel, Lisbon, Portugal, 
Septtrmber 1-S) - 

PitOlado, J.R. I''e7a. Lake Water QualitjY Honi'.orina Based on 
Rs mptejy Sensed Data: FinaJ Repprt . MOE'ROC Project No. 267RR. 
(Unpubl ished report > . 

Pitblado, J. P. 1997D. Remote sensing determinations o* Northeastern 
Ontario lake characteristics. Proceedings . Part B. Water Qui^l j^y 
Re5a4r£j3, 3 pp., Technoloigy Transfer Conference, Ontario Ministry 
of the tnvi ronmen t , Rcval York Hotel , Toronto, 'Jovewoer 30 
- Dee«ff>Ber I , 

Pitblado. J. Roger. 1<?BB. Differences in Lancsat MSS'TII sen^^or 
responses for acidic and non-acidic lake* in the SiidPury area. 
31 Bt Conference, International A»soc lat lon for jreat Lakes Research. 
McMastor University , Hami It on, Ontario. May 17-:!0. 

f*ii; bards, J. A. l?Sb. Romote Synsing Digital Image ftnalvsis . 
N. y . : Sor ingei — Ver lag. 

Scar pace, F,L., Holmqul^t, K.W.. and Fisher, L.'. 1979. 

Landsat analy«i« of lake quality. Photogrammetri^ , ,^n, g^neer jnq 
#nd Rf!mo te Sensing aS: 623-633. 

SPSS Inc. 19BH, 5PS5-K User ■ a Gmde - 3rd Edition, Chicago: 
SPSS Inc. 

Verdin. J. P. 19B5. Monitoring water quality conditions in a 

large western reservoir with Landsat imagery. Fhotoorammetnc 
^noin^erjnq and Remote Sensing 51:343-353. 

W^tzig, A. and Whilehurst, C. 19B1 . Literature review of the 

current use and technology of MSS digital data »or lake trophic 
classification. Proceedino^ Qf the I'JBl Pall Weet>no _ pf ^he 
American Society of Photooi'a'nmetrv . San Franciscci. pp. 1-20. 



Relationship of Mercury Levels in SportEish with Lake 
Sediment and Water Quality Variables. 

Christopher wren, B.A.R. Environmental, Nicholas Beaver Park, 
R.R. (13, Guelph, Ontario, NIH 6H9 (519) 824-5000 


Tissue mercury levels in smallroouth bass and walleye were weakly 
correlated with background sediment mercury levels. Mercury 
levels in lake trout were not correlated with background sediment 
mercury concentrations. 

Results suggest that geological mercury levels do not account for 
differences in fish mercury levels observed between lakes. 
Mercury concentrations in standard size smallmouth bass (31 cm) 
and walleye (41 cm) were negatively correlated with water quality 
variables reflecting water hardness and acidity. Therefore, fish 
of these species in low pH lakes tend to have elevated mercury 
levels relative to circumneutral lakes. Mercury concentrations in 
standard size <44 cm) lake trout were positively correlated with 
dissolved organic carbon and lake area. 

There was a very high correlation between standardized mercury 
concentrations In smallmouth bass and walleye, and between 
smallmouth bass and lake trout. The development of interspecies 
correlations could provide a useful management tool. 

The results of this study support the premise that a number of 
lake physical, chemical and biological variables simultaneously 
influence mercury accumulation and availability within lakes. 



The primary objective of this study was to examine the 
relationship between mercury levels in sport fish and natural 
sediment mercury content of lakes. 


It is well established that mercury levels in fish even from 
waters remote from direct pollution vary significantly between 
lakes, but the overall factors determining spatial variability 
remain obsure. A number of studies have reported that fish 
mercury levels are elevated in low pH lakes, suggesting that lake 
acidification is a factor affecting mercury uptake in fish 
{BrouESB et al 1977 : Suns et al 1987 : Wren and MacCrimmon 1983 : 
Bjorklund et al laB'i) . A variety of other biotic and abiotic 
variables are also known to influence mercury uptake in fish (eg. 
Richaan et al 1988; VerCa 1984; 1985). 

Mercury occurs naturally in combination with sulphide (HgS) known 
as cinnibar (Boyle 1974). The mercury content of ores generally 
increases with increasing zinc content, indicating that most 
mercury is present as a constituent of sphalerite (ZnS) , and is 
much less abundant in copper concentrates (Maclatchy and Jonasson 
1974) . 

Bedrock geology and soil type are known to have a profound 
influence on the chemical composition of many plant species, 
which in turn can directly affect the levels of certain elements 
(e.g. Se, cu. Mo) in terrestrial animals. However, the 
relationship between bedrock geology and mercury burdens in fish 
has not been seriously investigated. 

Some preliminary comparisons of fish mercury levels with sediment 
mercury levels have failed to establish a simple relationship 
between these two variables (e.g. McFarlane and Franz in 1980) . 
However, many of those studies were based on a relatively smal 1 
number of lakes. Bjorklund et al, (1984) reported a relationship 
between fish mercury burdens and surface sediment levels in 
Sweden. Hakonson (1980) developed a simple model to predict the 
mercury content of pike in Sweden based on surface sediment 
mercury content, water pH and a lake productivity index. 

Detailed lake sediment geochemical data, including mercury, are 
available for areas of Ontario through the Geological Survey of 
Canada. These data were used to investigate the potential role of 
natural geological mercury levels in affecting mercury levels in 
fish. Shi Its (1982) states that metal enrichment {in biota or 
sediments) in certain areas cannot be casually attributed to 
man's activities without knowledge of bedrock and sediment 



3.1 Data Col lectipr^ 

3.1.1 Lake Sediment Geochemistry 

Lake sedinent geochenical data were obtained from the GSC 
{Geological Survey of Canada) , Energy Mines and Resources in 

The GSC data were collected as part of reconnaissance surveys 
designed to gather a single index sample from a large number of 
lakes over a wide area. Both sediment and water samples are 
collected from each lake visited. Lake sediment samples were 
collected from a helicopter during the open water period. 
Samples were collected from the deepest portion of the lake. The 
theory is that a midlake sample is the most representative of a 
watershed erosional product. 

3.1.2 Provincial Water Quality and Fish Mercury Data 

The Ontario Ministry of the Environment and the Ontario Ministry 
of Natural Resources have jointly developed databases 
incorporating mercury levels at a standard fish length, lake 
physical characteristics and water quality data for a number of 
lakes in Ontario. Databases were available for smallmouth bass, 
lake trout, and walleye. Mercury levels at standard length were 
available for the following number oE lakes: Smallmouth bass, 
91f Lake trout, 91; Walleye , 255. 

All mercury concentrations in fish tissue were analyzed by the 
Ontario Ministry of the Environment (OME 1981). 

Individual lakes were chosen on the basis that at least 10 fish 
were sampled for mercury from that lake, and that there was a 
statistically significant correlation {p < 0.05) between log 
mercury and total fish length {McHurtry 1936). To compare mercury 
levels between lakes without a length bias, the average mercury 
concentration was predicted for a hypothetical fish of a standard 
length. The following standard lengths were utilized for the 
three species : Sma llmouth bass , 31 cm; Lake trout , Hi cm ; and 
Walleye, 41 cm. 

Water guality data from the Provincial Acid Sensitivity database 
were incorporated with the fish mercury data set. Sediment 
geochemical data were merged with the water chemistry and fish 
mercury data for corresponding lakes. 


3.2 Data Analysis 

Summary statistics, Pearson correlation coefficients, residuals 
and stepwise multiple regression analysis were conducted using 

The logjo of standardized fish mercury was used as the 
independent variable for all regression analysis. Some water 
quality variables were transformed for regression analysis as 
described by McMurtry (1986) . Sediment mercury concentrations 
were normal so the data were not transformed. Subsets of lake 
variables were entered as independent variables in stepwise 
regression analysis. Entering subsets of independent variables 
reduced two fundamental problems in multiple regression analysis: 
a) multicolinearity, and b) missing values. 


4.1 Smallmo uth; ba^S 

The mean standarized fish mercury concentration for a 31 cm bass 
was 402 ng/g (range 132-943 ng/g) . The mean pH of these lakes was 
6.97 (range 5.6-8.2). The mean alkalinity was 33.9 mg/L. The 
mean sediment mercury concentration was 99 ng/g (range 5- ISO 
ng/g) . 

The standardized log fish mercury concentration was significantly 
correlated with a number of sediment and water quality variables. 
Fish mercury was positively correlated with sediment mercury 
concentrations (r = 0.31, Figure 1), and negatively correlated 
with lake pH (r - -0.52; Figure 2). 

The mean mercury concentration in smallmouth bass in lakes with 
pH i 6.5 was 353 ng/g (n = 49), compared with 545 ng/g in lakes 
with pH < 6.5 (n " 17) . 

4.2 Lake Trout 

The mean standardized mercury concentration in lake trout was 303 
ng/g (n = 41 lakes, range ■« 70 - 1033 ng/g). The mean sediment 
mercury concentration in lake trout lakes was 138 ng/g. 

The mean pH of the lake trout lakes was 6,4, The average pH of 
lake trout lakes was substantially lower than the average pH of 
either smallmouth bass or walleye lakes. 


Fish mercury concentration was not significantly correlated 
(p > 0.05) with lake pH (r = 0. 34) or sediment mercury 
concentration (r = - 0, 10) . The relationship between mercury 
concentration In lake trout and sediment mercury concentration is 
illustrated in Figure 3. 

The mean standardized mercury concentration in walleye was 517 
ng/g (n= 44 lakes, range = 12B-2216) . The mean sediment mercury 
concentration was 88 ng/g (range 10-18B) . 

The average pH of the walleye lakes was 7.30. The average pH was 
similar to smallmouth bass lakes but the range was not as great. 
For example, only 2 of the 44 walleye lakes had pH < 6.5. 

Stepwise multiple linear regression analysis consistently chose 
conductivity as the single best predictor of log fish mercury 
concentration . 

4.4 Interspecies Correlations 

There was a good correlation of standard mercury concentrations 
between species among overlapping lakes. Figures 4 illustrates 
the relationships between smallmouth bass and walleye mercury 
concentrations . 


The results suggest that background sediment mercury levels were 
significantly correlated with mercury levels in smallmouth bass 
and walleye. However, the background sediment mercury levels did 
not explain the differences observed in fish mercury levels 
between lakes. 

The average background sediment mercury levels in this study 
agree very closely with other studies that suggest the background 
mercury concentration in lake sediments is approximately 100 ng/g 
(Forstner and Whittman 1981; Cahill and Shimp 1984; Bjorklund et 
al 1984; Evans 1986) . 

The results of this study and those of Johnson ( 1987 ) and 
Bjorklund ( 1984 ) suggest that surface sediment mercury 
concentrations may be better than background sediment levels as 
an indicator of mercury availability, and fish mercury levels 
within a lake. However, since other factors such as water quality 
have an obvious influence on fish mercury levels, it is becoming 
apparent that fish mercury levels cannot be accurately predicted 
by a single environmental variable. 


Mercury levels in smallmouth bass and walleye were highly 
correlated with variables reflecting water hardness and acidity. 
Calcium and conductivity were generally better single predictors 
of fish mercury than lake pH. This may be partially attributed to 
the quality and variability of lake pH data, and also to greater 
direct influence of calcium than pK on mercury uptake. 

Water hardness and lake pH likely influence mercury uptake and 
availability simultaneously within a lake. Rodgers and Beamish 
(198 3) reported that the efficiency of methyl mercury uptake by 
rainbow trout was much greater in sof twater (30 mg/1 as caC03) 
compared with hardwater (385 mg/1 as CaC03) . The mechanisms to 
account for this effect may be increased gill permeability at low 
Ca levels (Spry et al 1981) or competition between metals and Ca 
for cellular binding sites (Zitko and Carson 1976). 

Reduced lake pH may increase the bioavailability of mercury by 
stimulating bacterial methylation from the sediments (Xun et al 
1987) , Thus, a combination of increased production and uptake 
efficiency could eicplain elevated mercury levels in fish from low 
pH lakes. 

Negative correlations between lake pH and/ or alkalinity and 
mercury content of fish in Ontario have now been demonstrated for 
yearling yellow perch (Suns et al 1980) , pumpkinseed sunf ish 
(Wren and HacCrimmon 1983), smallmouth bass (McHurtry 1987; Suns 
et al 1987) and now walleye (this study) . Of these, the walleye 
lakes had the greatest geographical distribution in the province. 

Mercury concentrations in standard length lake trout were highly 
correlated to DOC and lake area, but not water hardness or 
acidity variables. Dissolved organic carbon was also selected in 
conjunction with calcium as predictors of mercury levels in 
smallmouth bass. Studies in Sweden and Finland have also noted a 
positive correlation between fish mercury levels and the humic 
content of water (Verta 1984; 1985; Lindqvist et al 19B6) . It is 
suggested that the humic substances act as both an energy source 
and source of mercury for the methylating bacteria (Verta 1985). 
Bodaly and Hecky (1984) suggest that increased availability of 
organic material accounts for elevated fish mercury levels in new 
hydroelectric reservoirs. 

It is notable that the average mercury concentration in a 41 cm 
walleye (517 ng/g) is above the recommended safe level of mercury 
in fish for unlimited consumption, A length of 41 cm corresponds 
to the average length of walleye sampled in the sportfish 
contaminant monitoring program (Scheider pers commun) . 
The demonstrated relationship of mercury concentrations between 
species is significant- This is the first study to document a 
relationship between mercury levels in sportfish from such a 
large number of lakes. A relationship between mercury levels in 


different fish species has great potential as a management tool 
both for predicting mercury levels in other species, and for 
designing fish contaminant monitoring programs. 

The results indicate that background sediment mercury levels do 
not account for the observed differences in mercury levels in 
fish between lakes. The differences may often be explained by 
water quality, expecially DOC, pH and alkalinity. Therefore, the 
factors and conditions affecting these variables in Ontaro lakes 
will also influence mercury availability and uptake in fish. 
These variables will act simultaneously on mercury uptake and 
availability, and the net effect from individual variables will 
differ between species and lakes. 


Bjorklund, I., H. Borg and K. Johansson. 1984. Mercury in Swedish 
lakes - its regional distribution and causes. Ambio 
13(2) :118-121. 

Bodaly, R.A., R.E. Hecky and R.J. Fudge, 1984- Increases in fish 
mercury levels in lakes flooded by the Churchill River 
diversion , northern Manitoba . Can. J . Fish . Aquat . Sci . 

Boyle, R.W. 1974. Elemental associations in mineral deposits and 
indicator elements of interest in geochemical prospecting. 
Geol. Surv. of Canada Paper 74-45. 

Brouzes, R.J. , R.A. Mclean and G.H, Tomlinson. 1977. The link 
between pH of natural waters and the mercury content of 
f ish . Research Report to the U.S. National Academy of 
Sciences, Dontar Res. Rep. , Senneville, Que. 

Cahill, R.A, and N.F. Shimp. 1984. Inorganic contaminants in Lake 
Michigan sediments, in J- Nriagu and M. Simmons (edaj Toxic 
substances in the Great Lakes. J. Wiley and Sons, New York p394 . 

Coker, W.B. and I. Nichol. 1975. The relation of lake sediment 
geochemistry to mineralization in the northwest Ontario 
region of the Canadian shield. Economic Geol. 70:202-218. 

Evans, R.D. 1986. Sources of mercury contamination in the 
sediments of small headwater lakes in south-central Ontario, 
Canada. Arch. Envir. Contam. Toxicol. 15:505-512. 

Forstner, u. and G.T, Whittmann. 1981. Metal pollution in the 
aquatic environment. Springer-Vewrlag. Berlin. 


Hakonson, L. 1980. The quantitative Impact of pH, bioproduction 
and Hg-contamination on the Hg-content oC fish (pike) , 
Envir. Poltut. (Series BjiasS-SOi. 

Johnson, M.G. , L.R. Gulp and S.E. Gaorga. 1986, temporal and 
spatial trends in metal loadings to sediments of the Turkey 
Lakes, Ontario. J. Can. Fish. Aquat. Sci. 43:751-762. 

Johnson, M.G. 1987. Trace element loading to sediments of 
fourteen Ontario lakes and correlations with concentrations 
in fish. can. J. Fish. Aq. Sci. 44:3-13. 

Maclatchy, J.E. and I.R. Jonasson. 1974. The relationship between 
mercury occurrence and mining activity in the Nottaway and 
Rupert River basins of northwestern Quebec. Geo!. Survey o£ 
Canada GSC Paper 74-56. 

McFarlane, G.A. and W.G. Franzin. 19B0. An examination of Cd, 
cu, and Hg concentrations in livers of northern pike and 
white sucker from five lakes near a base metal smelter at 
Flin Floon, Manitoba. Can. J. Fish. Aquat. Sci. 37: 1573- 

McKurtry, M. 1986. The relationship between mercury levels in 
lake trout and water quality parameters, unpublshed report, 
Ont. Win. Environ. 

Richman, L. , P.M. Stokes and CD. Wren. 1988. A review of facts 
and fallacies concerning mercury uptake in fish from acid 
stressed lakes. Water, Air Soil Pollut. (in press, accepted 
November 1987) . 

Rodger, D. and F. W. Beamish. 1983. Water quality modifies uptake 
of woterborne methylmercury by rainbow trout. Can. J. Fish. 
Aquat. Sci. 40:824-828. 

Shilts, W.W. 1982. Potential effects of acid rain on glaciated 
terrain. In: R.G. LaFluer (ed) Groundwater as a geomorphic 
agent. Allen and Unwin Inc. Boston. 

Suns, K., Hitchin, G. , Loescher, B. , Pastorek, E. and Pearce, R. 
19B7. Metal accumulation in fishes from Huskoka-Haliburton 
lakes in Ontario (1978-1984). Ont. Min. Envir, Tech. Rep. 

Suns, K., C. Curry and D. Russell. 1980. The effects of water 
quality and tnorphometric parameters on mercury uptake by 


yearling yellow perch. Ont. Min. Env. Tech. Rep. LTS BO-1, 

Spry, D.G. , CM. wood and P.V. Hodson. 1981. The effects of 
environmental acid on freshwater fish with particular 
reference to the softwater lakes of Ontario and the 
modifying effects of heavy metals. Can. tech. Rep. Fish. 
Aguat. Sci. No 999. 

Verta, M. 1985. The effect of water quality on mercury 
accumulation in polyhumic lakes and reservoirs. In, Proc. 
Int. Conf. Heavy Metals in the Environ., Vol. 1, Athens, 
Sept. 19aB. pp 103-106. 

Wren, CD. and H.R. MacCrimmon. 1983. Mercury levels in sunfish 
relative to lake pH and other environmental variables of 
Precambrian shield lakes. Can. J. Fish. Aquat. Sci. 40:1737- 

Xun, L. , Campbell, N.E. and Rudd, J.W. 1987. Measurements of 
specific rates of net methylmercury production in the water 
column and surface sediments of acidified and circumneutal 
lakes. Can. J. Fish. Aquat. Sci. 44:750-757. 

Zitko, V. and W. Carson. 1976. A mechanism of the effects of 
water hardness on the lethality of heavy metals to fish. 
Chemosphere 5:299-303. 



SmallniouLb Bais Lakes 

>a M IM lis 

ri4ura I. tha retitlanlhlp brtartn it 4tiiJ*rd If b« ncroury Ivvrl-. 
In koAllaouth batt t"6 I aKd iflillmcnt ncrcury Invrl, 






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In V'^BlM^hfl'- I-^bI r«P&) 

rcljlliin [n It jnddrdliC'i] wrcury runcent r.i' la 
OHri-inoiitti lidii III (IdlHriD Idkfi. 





A. Ian HcLeod 

Dapartinent of Statistical and Actuarial Science 

The University of Western Ontario, London, Ontario 

Keith W, Hlpe: 
Depai-tments of Systens Deaign Engineering 

and statistics and Actuarial Science 
Untverolty of Waterloo, Waterloo, Ontario 

Byron Bodo 

Water Pesourcea Branch, Ontario Ministry of the Environment 

and Adjunct Professor In the Department of 

Statistical and Actuarial Sciences 

The Unireralty of Western Ontario 


The overall objec^tlve of the study Is to develop graphical and statistical 
trend analysis procedures for use with the water quality time series obtained 
from Ontario's Provincial Water Quality Monitoring Network (PWOHN). Trend 
analysis is required for alerting authorities about «iter quality degradation 
so that appropriate corrective action can be taken and evaluating the perfor- 
mance of pollution abatement sohemes. In order to detect visually Increasing 
or decreasing trends, a range of exploratory data analysts techniques are 
being applied to PWQMN data sets. The graphical methods Include time series 
plots. bo» and whisker graphs and a SBOothlng teohnlque called robust locally 
weighted regression. Special types of nonparametrlc tests are being refined 
and developed for rigorously testing for the presence of trends. These tests 
will also be applied to PWQKN water quality time series. 



Water quality and other kinds of environmental time aecleg often poaaeaa 
oKaractBrlatlca which do not allow the aeries to be easily analyzed using 
statistical techniques [3 to 7, 9, 11, 131. One of the major problema with 
these time series I.s that there are often many missing data points among which 
there may be long periods of time for which no observations wore taken. Water 
quality data may be non-normally distributed and follow a distribution i^ich 
la usually positively skewed. In addition, the data are often censored by 
only listing measiiremonta below a certain level as being "less than" or measu- 
rements above a specified level as being "greater than". For instance, con- 
centration values for toxic compounds, metals or organic compounds which fall 
below the lljnlts of detection for certain chemical testa are reported simply 
as less than the lluills of detection. Seasonality effects contained in a 
aequence of measurements over the years can cause cyclic patterns to appear 
In a graph of a given water quality variable. When there are many different 
water quality variables Interacting with one another in the presence of vary- 
ing flow rates, the roultlvarlate effects of the Interactions can be quite 
complex. As a further major ooaiplioatlon, cme or more external Interventions 
may slgnlfloantly affect the stochastic manner in which a aeries behaves and 
thereby create a variety of trends. Because of the foregoing and other 
reasons, environmental data are often quite "messy". 

In order to extract an optimal amount of information from messy environ- 
mental data, a systems design approach to data analysis can bo followed. As 
proposed by Tukey [It] and demonstrated by authors such as McLeod et al. [11] 
and HIpel et al. [5l using water quality data, the two major steps In a 
statistical study oonslst of exploratory and confirmatory data analysis. The 
objective of the exploratory data analysis stage is to employ simple graphical 
and numerleal techniques to discover Important patterns and statistical char- 
acteristics such as the presence of trends. The purposes of the confirmatory 
data analysis stage are to oonflrm statistically In a rigorous fashion the 
presence or absence of certain properties in the data. Depending upon the 
quantity and quality of the data being analyzed, appropriate graphical, para- 
netrlc and aonparametrlc techniques can be employed as exploratory and confir- 
matory data analysis tools, 

Both exploratory and confirmatory data analysis methods arc used In this 


study Tor d»t«cMog and modelling trends contained in water quality tlae 
series obtained frem Ontario's Provincial Water Quality Monitoring Network 
(PWOhTN). In the next section, representative results are presented for demon- 
strating how some different ielnda of graphs can be used as exploratory tools 
for visually detecting trends and other statistical properties in water qual- 
ity series. Following this, It Is explained how a number of nonparametrlc 
tests are being developed and enployetl for finding trends In messy water 
quality time aeries. 


Different types of grains are available for use as exploratory data analy- 
sis tools for detecting trends In a data set. Table 1 lists some of those 
graphs along with brief descriptions of their purposes and references contain- 
ing detailed explanations about the technlctues . Applications of sone of these 
graphical methods to water quality time aeries are presented in publications 
by authors such as El-Shasrawi and Kwlatowaki [3], MoLeod et al. [11], Hlpel 
et al. [5] and Hlpel [U], Below, representative results are given for the 
cases when two types of the graphs In Table 1 are used with a PWQMN data set. 

Table 1. 
Graphs for use In Trend Analysis at the Exploratory Data Analysis Stage 

Types of Graphs Purposes 

Time Series Plot 

detect trends and other 
statistical characteristics. 

References for Desorlptioos 

Tukey [lU]. Moat 
statistical textbooks. 

Box and Whisker 

Star Symbol 

Tukey Blurred 

Robust Locally 

Regression Smooth 

Graphically oummarlze Import- Tukey t1't| Ch. 2] 
ant statistics of data for HoGlU at al. [10] 
each season of the year. 
Compare plots before and 
after the Intervention. 

Display how medians and other Chambers et al. [1] 
statistics change across 
seasons and years. 

Trace trends 

Trace the general shape of 
trends In a time series plot 

Tukey [11, Ch. 7] 
McNeil [12] 

Cleveland [2] 


Removal of the Effeota of Flow 

The statistical properties of a given water quality variable are often 
dependent upon rlverflowa, as well aa other physical feotors. After removing 
the statistical effects of flows contained In the water quality time series, 
one can check for the presanoe of trends In the residual or filtered water 
quality time sorles. One approach for removing flow levels, represented by 
xi, from the water quality variable, y^, la to employ the cubic regression 
analysis equation given by 

Vi = a + Bj, Xj + B2 x^-^ + Bj x^3 * B^ (!)■ 

-ih ere a la the level paraneter; Bj, J= 1,2, 3i are the regression parameters; 
and ei represents the residual series. Subsequently, exploratory and conflr- 
aatory data analysis tools ean be used for detecting and modelling trends In 
the residual aeries in (1). Furthermore, irtien deemed necessary, one may wish 
to take natural logarithms or some other kind of data transformation of the yi 
and/or ici aeries before performing the regreaalon analysis. 

On the Grand River at Dunnvllle, Ontario, there are 1700 average dally 
values of total phosphorous (mg/i) available from 1972 to 1985, as well as 
average dally riverflows (mS/s). When equation (D Is used to regress the 
logaflthiuio total phosphorous observations upon the logarithmic flows, all of 
tne regression paraoeters are significantly different from zero. 
Consequently, in the applications the residuals of the regression analysis are 

Box and Wh laker Graphs 

A box and whisker graph la based upon kriiat la called a ^-number summary 
Lia, Ch, 2]. For a given data set, the 5-number sunanary oonslsts of the smal- 
lest and largest values, the median, and the 0.25 and 0.75 quantlles, which 
are called hingea. When the data are ranked frrao largest to smallest, the 
first data point is the smallest value while the last entry is the largest. 
When examining a seasonal time series, such as monthly or quarterly data, it 
la Inatruotlve to calculate a 5-number summary plus certain types of extreme 
values for each season. A convenient manner in which to display this Informa- 
tion is to plot a "box and whisker" diagram for each season or month. 

Figure 1 displays a notched box and whisker plot for each month of the 
residuals for the total phosphorous series at Dunvllle, Ontario. For a given 


month, the uppar and lower ends of a notched rectangle reprasent the two 

hinges and the thick line dr-awn horizontally in the rectangle Is the value of 
the median. The width of each box la proportlcmfll to the number of data 
points in the corresponding seasmi. Excluding extr«ie values, the minimum and 
BaxlinuiD values for a particular rnwith are the end points of the lines or 
"whiskers" attached to the rectangle or box. A special type of extreme value 
called a far-out value [11, Ch. 2] is Indicated by the points plotted above 
and below the whiskers In a given season. The notch on both sides of the 
median allows one to compare medians across months or seasons. If, when 
comparing two months, the notches overlap, one can argue that the medians for 
those months are not significantly different from one another at the 95J 
confidence level. In subsequent analyses, one may wish to join similar months 
together aa a single season In order to provide results which are simple and 
easier to understand. 

Figure 2 shows a graph of the average residual aeries for total phosphor- 
ous when the 12 monthly series are oranblned as five saasonal residual aeries. 
The legend explains how consecutive months are Joined together. For example, 
MDJ means the residuals for November, December and January are considered as a 
single season across all the years. The five time series plots in Figure 2 
for the total phosphorous residuals, clearly portray a downward trend In the 
earlier 1970'a which then levels off from the nid-1970's onwards. 

Honparaitetrlc Tests 

Various kinds of graphs are used as exploratory data analysis tools for 
visually finding trends in a time series. At the confirmatory data analysis 
stage, atatiatlcal tests can be employed for detecting trends and perhaps also 
estimating their magnitudes. In order to lessen the number of underlying 
assumptions required for testing a hypothesis such as the presence of a speci- 
fic kind of trend in a data set, researchers developed nonparametric testa 
[8]. Nonparametric tests were developed for use in environmental impact 
assessment because scientists were concerned that the statistical characteris- 
tics of messy environmental data would malte It difficult to use the parametric 
procedures [3 to 7. 9t 13]. 

In this study, a number of nonpar a met rlo teats are being refined for ap- 
plication to PWQMN water quality time series. One parametric trend test that 
l8 currently being applied to the data Is a version of the seasonal Mann- 


Kendall teat [5,7,131. Kendall's partial rank correlation test [3] la also 
being further developed so it can be applied to water quality time series. 
Instead of using a paranetric procedure such as the regression analysis method 
In (I) to remove flow effects ir a water quality series, one can employ the 
partial rank correlation approach to both partial out the flows and test for 
the presence of a nonotonlc trend over tine in the water quality series being 


PI Chambers, J.M., Cleveland, W.S., Kleiner, 8., and Tukey, P. A., "Graphical 
Methods for Data Analysis", Duxbury Press, Boston, 1983. 

t2l Cleveland, W.S., "Robust Locally Weighted Hegresslon and Smoothing 
Scatterplots", Journal of the American Statistical Association, Vol. TUi 
No. 368, pp. 829-836, 1979. 

[3] El-Shaarawl, A.H., and Kwlatowskl, R.E. (Editors), "Statistical Aspects 
of Water Quality Monitoring", Elsevier, Amsterdam, 1985. 

CI Hlpel, K.W. (Editor), "Nonparametrlc Approaches to Envlronaenlal Impact 
Assessment", Monograph published by the American Water Resources Associa- 
tion, iimo Grosvenor Une, Suite 2^0, Bethesda, Maryland, U.S.A., 1996. 

C5] Hlpel, K.W., McLeod, A.I. , and Weiler, R.B., "Data Analysis of Hater 
Quality Time Series In Lake Erie", Water Resources Bulletin, Vol. 21, No. 
3, pp. 533-5')'', 1988. 

[6] Rlrsch, R.M. and Slafjk,, "A Nonparametrlc Trend Test for Seasonal 
Data with Serial Dependence", Water Resources Research, Vol. 20, (Jo. 6, 
pp. 727-732. 198«. 

[7] Hlrach, R.M., Slack, J.R., and Smith, R.A., "Techniques for Trend Assess- 
ment for Monthly Water Quality Data", Water aeaouroea Research, Vol. 18, 
No. 1, pp. 107-121, 1962. 

[S] Kendall, M.G., "Rank Correlation Methods", Charles Griffin, London, 
Fourth Edition, 1975. 

[9] l.ettenmaler, O.P., "Detection of Trends in Water Quality Data from Reco- 
rds with Dependent Obaervatlons", Water Resources Research, Vol. 12, No. 
5, pp. 1037-1016, 1976. 

[10] HcGlll, R., Tukey, J.W. and Uren, W.A., "Variation of Boat Plots", 
American Statistician, Vol. 32, pp. 2-12, 1983. 

[11] McLeod, A.I., Hlpel, K.W., and Camaoho, F., "Trend Aaaesament of Water 
Quality Time Series", Water Resources Bulletin, Vol. 19, No. t, pp. 537- 
517, 1983. 

[12] McNeil, D.R., "Interactive Data Analysis", Wiley, New York, 586 pp., 

[13] Van Belle, G. and Hughes, J. P., "Nonparametrlc Tests for Trend in Water 
Quality", Water Resources Research, Vol. 20, No. 1, pp. 127-136, 1981. 

[11] Tukey, J., "Exploratory Data Analysis", Addlson-Wesley, Reading, Mass., 



A a as 9 9 jj J 

J L 

3an F»h fWr- Bpr 

nmu Jun All *•« (in 0=t —^ <>" 


Figute I. Box and Whisker Plots for the ReaidualB of the Total PhosphorouH MeasuireBHiita on 
the Grand River at Dunnville, Ontario. 


Figure 3. Seasonal Residual Series of the Total PhWH^iboroua OxamrvaZions cm the Grand 
Rlvec at Dimnvllla, Ontario 


Use of a Broaobenzoate for Cross-adaptation of Anaerobic Bacteria 
In l^ke Ontario Sediments for Degradation of Chlorinated Aromatics 

Martina Urfaanek^ Tatiana strycek^ Campbell Wyndham^ and 
Morris Goldner^ 

^Department of Microbiology, University of Toronto, Toronto, 
2 Ontario, Canada nss 1A8 

Department of Biology and Environmental Laboratories, Carleton 
University, Ottawa, Ontario KIS 5B6 


In surveys of Toronto waterfront sediments, Persaud et al. 
{1995) disclosed that muds in boat slips of the Inner Harbour and 
in Toronto Island waterways had elevated levels of total volatile 
residues, solvent extractables, oil and grease, and PCB's. Many such 
compounds were originally believed to be recalcitrant. Within the 
past few years several research groups have found that chlorinated 
aromatic compounds can be decomposed in the laboratory by 
stringently anaerobic bacterial consortia present in lake sediments 
(Horowitz et al. 1982, 1983; Suflita et al . 1982, 19S3; Shelton & 
Tied:)e l9S4a, 1984b; Tiedje et al. 1986). 

In the light of these findings, studies were undertaken in 
order to investigate the potential of sediment microorganisms for 
anaerobic degradation of halogenated pollutants. Sediments were 
collected from sites along the Toronto waterfront in Lake Ontario 
(Toronto, Ontario, Canada). These were anaerobically incubated with 
monohalogenated benzoates. Following adaptation, as measured by the 
rate of substrate disappearance, the sediments were subsequently 
incubated with polysubstituted benzoates. Cross-adaptation to these 
complex aromatics was assessed by high pressure liquid 
chromatography (HPLC) . The findings demonstrate a potential for 
breakdown of halogenated contaminants by anaerobic microorganisms 
found in Lake Ontario sediment. This points to the feasibility of 
applications of preadapted microbial consortia for anaerobic 
degradation of xenobiotlcs under controlled conditions- 


Sediment collection 

sediment samples were collected in Lake Ontario from sites located 
along the Toronto waterfront by Ponar grab sampler, and kept in 
glass jars (0.5 1 capacity) at 4°C in the dark under anaerobic 
conditions until further use. The sampling sites were characterized 
by organic carbon-rich sediments which were anaerobic at all depths. 

preparation of test medium 

Revised Anaerobic Mineral Medium (RAHM) was used as suspending 

medium (Shelton (. Tiedje, 1984a). The medium contained phosphate 
buffer (pH 7.0), mineral salts, trace metals, vitamins, reducinq 
agent (0.125% L-cysteine . HCl -Na.S. 9H-,0, pH 10}, and a redox 


indicator (0. 1% resazurin) . Preparation and transfer of media and 
inocula were carried out in an anaerobic hood with protective 
eyewear and gloves, Hungate technique and apparatus (Hungate, 1968) , 
were used for anaerobic gassing of flasks and for preparation of 
oxygen-tree gases and media. 

Chein icai.5 

3-broinobenzoace {3-BrBZ) and 3-chlorobenzoate (3-C1B2) were obtained 
from Sigma Chemical Co., St. Louis, MO.; 3 , 5-dichlorobenzoate (3,5- 
diClBZ) was supplied by Fluka Chemical Corp. , Ronkonkoma , NY . ; and, 
4-ainino-3 , 5-dichlorobenzoate (-l-anino-S, 5-diClBZ) was obtained from 
Pfaltz and Bauer Research Chemical Division, Waterbury, CT. 

Water insoluble test compounds were first dissolved in IM NaOH. 
Appropriate amounts were dispensed through a disposable membrane 
filter (0.45 urn, Hillipore Ltd., Montreal, Que.). 

Cross-adaptation eyperijuent 

Collected sediment samples in volumes of 500 nL were anaeroblcally 
transferred to 2-1 Erlenmeyer flasks and then suspended in RAMM in a 
1:2 ratio. The flasks were sealed with thick butyl rubber stoppers. 
The stoppers were secured in place with tape and equipped with a 
needle attached to Nalgene (Nalge Co. , Division of sybron Corp. , 
Rochester, NV.) tubing trapped in a test tube filled with water to 
maintain atmospheric pressure in the head space of the flasks. The 
substrates, 3-ClBZ and 3-BrBZ, were added to the sediment slurry in 
the flasks at concentrations in the range of 0.6 - 1.2 mM. 
Incubation was carried out in the dark at room temperature. Samples 
of sediment pore water were taken from flasks at regular time 
intervals, passed through a 0.45um Hillipore disposable membrane 
filter, and stored at -10° c for HPLC analysis. Substrate depletion 
was monitored in a Waters Hillipore system (Waters Division of 
Hillipore (Canada) Ltd., Mississauga, Ont., Waters 501 HPLC pump, 
Novapak C^g column, 50% methanol/ 1% acetic acid as solvents , UV 
absorbance detection at 2 54 nm, 10 ul sample loop) . Once total 
substrate depletion occurred, the same substrate (0.6 - 1.2 mM) was 
added again. The sediment consortium was considered to be adapted 
after a repeated addition showed the substrate to be degraded 
without a lag period or with a much shortened lag period and with a 
noticeably diminished time for its complete depletion. 

For cross-adaptation experiments, the sediments previously 
adapted to a monohalogenated substrate were incubated with a 
polyhalogenated substrate (0.6 - 1.2 mM) , 3,5-diClBZ or 4-amino-3,5- 
diClBZ. Autoc laved (30 min, 10 Ibs/sq. in. ) sediments served as 
controls and were incubated under the same conditions as test 
flasks, with substrate concentrations monitored by HPLC 


The ability of sediment microorganisms to adapt and cross-adapt to 
halogenated compounds is naturally of great interest, considering 
the problem that these types of chemicals pose to our freshwater 
ecosystems. The cross -adaptation experiments provided some insight 
in this direction. 


HPLC analysis revealed that lag periods in non -adapted 
sediments were much shorter for 3-BrBZ than for 3-clBZ; but, once 

degradation commenced, it appeared to be guite rapid for both 
substrates (Fig. 1). Subsequent additions of these substrates to the 
same sediment resulted in a much shortened lag period (Fig. 1) while 
substrates were depleted in a similar pattern, both initially and 
following adaptation (Table 1) , 

Previous experiments (Strycek et al., 1987) involving the 
monitoring of gas production had revealed that both 3 , 5-diClBZ and 
4-amino-3 ,5-diClBZ were degraded very slowly, even after 12 weeks of 
incubation, with a minimal gas production, as compared to simpler 
aromatic compounds (e.q. 3-BrB2 or 1- iodobenzoate ) . However, 
organisms adapted to 3-ClBZ required four to five weeks for 3,5- 
diClBZ depletion. Organisms adapted to 3-BrBZ were able to 
completely deplete the 3,5-diClB2 in 3 weeks. The amino derivative, 
4-amino-3,5-diClBZ was only partially degraded by organisms adapted 
to 3-ClBZ even after 16 weeks; but, when the organisms were adapted 
to 3-BrBZ, they were able to completely degrade 4 -amino- 3, 5-diClBZ 
in six weeks (Figure 2, Table 2). 

The results from the different sediment sampling sites examined 
(six in total) were pooled together in order to provide 
representative data from which trends could be determined . The 
cross-adaptation showed that respectively 3,5-diClBZ and 4-amino- 
3 , 5-diClBZ were degraded more rapidly in sediments previously 
adapted to 3-BrBZ than in those adapted to 3-clBZ or in non-adapted 
ones. Thus, pre-adaptation to 3-BrBZ may be an effective measure in 
inducing breakdown of the polyhalogenated derivatives. 

The feasibility of inducing biologically mediated breakdown of 
complex halogsnated aroma tics suggests possible practical 
applications. A wider range of substrates should be tested in order 
to establish the generality of observations in this report. Given 
this further progress, adapted anaerobic sediment cultures might be 
usable either in in situ treatment of contaminated anaerobic 
sediments or in controlled wastewater treatment systems. 


The authors wish to express their great appreciation for Judith F.M. 
Hoeniqer's work in initiating and designing this project, and their 
deep sorrow in her loss. 


Horowitz, A. , D.R. Shelton, C.P. Cornell t Tiedje, J.H. 1982 
Anaerobic Degradation of Aromatic Compounds in Sediments and 
Digested Sludge. Dev. Indust. Microbiol . 23, 435-444 . 

Horowitz, A. , J.H. Suflita & Tiedje, J.H. 1983 Reductive 
Dehalogenation of Halobenzoates by Anaerobic Lake Sediment 
Microorganisms. Appl. Env. Microbiol. 45, 1459-1465. 

Hungate, R.E. 1968 A Roll Tube Method for cultivation of Strict 
Anaerobes. In Advances in microbiology ed. Norris, J.R. t, Ribbons, 
D.W. vol. 3B, pp. 117-132. New York: Academic Press Inc. 


Persaud, D. , T. Lomas, D. Boyd, & Hatha! , S. 1985 Historical 
development and quality of Toronto waterfront sediments. Part I. 
Ontario Ministry of Environment. 

Sheiton, D.R., & Tiedje, J.M. 1984a General Method for Determining 
Anaerobic Biodegradatlon Potential. Appl. Environ, Microbiol. 47, 

Shelton, D.R., & Tiedje, J.M. l9S4b Isolation and Partial 
Characterization of Bacteria in an Anaerobic Consortium That 
Mineralizes 3-Chlorobenzoic Acid. Appl ■ Environ, Microbiol . 48, 

Strycek. T. , Urbanek, K. , Hyndham, R. C. , Goldner, M. 1987 
Degradation of Organic Contaminants by Anaerobic Bacteria in Lake 
Ontario Sediments, Abstract B6. 1987 Environment Ontario Technology 
Transfer Conference Proceedings. 

Sufllta, J.M. , A. Horowitz, & Shelton, D.R. 1982 Dehalogenation: A 
Novel Pathway for the Anaerobic Biodegradation of Haloaromatic 
Compounds ■ Science. 218 , 111 5-1117 . 

Suflita, J.M. , J. A, Robinson & Tledje, J.M. 198 3 Kinetics of 
Microbial Dehalogenation of Haloaromatic Substrates In 
^5etha^ogenic Environments. Appl. Environ. Microbiol. 45, 1466-1473. 

Tiedje, J.H. , S.A. Boyd, & Fathepure, B.Z. 1986 Anaerobic 
Degradation of Chlorinated Aromatic Hydrocarbons. Dev. Ind. 
Microbiol . 27: 117-127 

List of Tables 

Table 1. Comparison of average times for substrate depletion in 
adapted and non-adapted sediments 

Table 2 . Average comparative times for substrate depletions in 
cross-adapted sediments 


Table 1. Conparison of average tiroes^ for substrate depletion in 
adapted and non-adapted sediments 

substrate 3-BrB2 3-ClBZ 

time required for 7 weeks 15 weeks 

initial substrate 


time required for 4 weeks 6 weeks 

subsequent substrate 


^Data pooled for six sampling sites on the Toronto Waterfront. 


Table 2. Average comparative tiroes^ for substrate depletions in 
cross-adapted sediments 

adapted to: 
cross-adapted to 



3 weeks 

6 weeks 


4 . 5 weeks 
16+ weeks 

*Data pooled for two (3,5-diClBZ) and tour [4-amino-3 , 5-diClB2) 
sampling sites on the Toronto waterfront. Sediments initially 
adapted to 3-BrBZ and 3-ClBZ. 

Legend for Figures 

Figure 1. Substrate depletion in sediment slurries from a typical 
sampling site on the Toronto waterfront. 

3-C1B2 breakdown in non-adapted sediments 

3-ClBZ breakdown in adapted sediments 

3-BrBZ breakdown in non-adapted sediments 

3-BrBZ breakdown in adapted sediments 

Figure 2. A typical pattern of cross-adaptation to polyhalogenated 
substrates. Compound in parenthesis indicates substrate used for 
initial adaptation. 



\ U8-W 














' '-\ 




„ 3-CI02\\ 




3-B.M \V 

\ ■ 

6 8 10 12 14 16 18 

Time (weeks) 



c 1.0 

~ 0.8 


5 0,6 - 

I 0.4h 

4-*mino-is.(uciaz a-cmz\ 

0.2 - \ 



i-i,s-oiC4oz la-aaii 

6 8 10 12 14 16 18 
T)m« (wMks) 




Poster Presentations 

'• ■ T 




Tha Effects of Agrlcult«r«l Dr&inaga 

S«dla«nc «tid Vacar Quality Loadlngt 

J.D. Paine, S.A. Facars. V.E. Uact 

DaparEinanc of Civil Engineering. 

Queen's Unlversl:y, Kingston, Ontario, K7L 3N6 

lapaimient of vacer quality as a result of agricultural activity is 
desired neither by the fanning comnunity nor by recreational users of 
receiving wac«r bodies . Removal, of valxiabla pesticides , fertilizers and 
copsoil by surface or subsurface drainaga represents an economic loss to 
agriculture and a potential pollution hazard to receiving waters. In order 
CO evaluate this potential iDpalroent of wacet quality, there Is a need for 
accurate prediction of chemical contsntlnant and sedinent loadings for 
different envlronnents. loading conditions , and agricultural nanagetvent 
strategies . Such an evaluation is best accomplished by a combination of 
selective measureinent and more general modelling. The aodel ecuployed should 
be deterministic and capable of simulating both water quantity and quality 
a: the field level. A few quantity nodels chat meet these criteria are 
available. However, because of the lack of applicable quality models, a 
major task in this project is the development of physically -based quality 
algorithms that can be linked to hydrologlc quantity models to accurately 
predict chaalcal contaminant and sediment loadings. 

An Ontario Ministry of the Environment funded research project 
[■ 152 FL> entitled The Effects of Tile Drainage and Open Ditches on Peak 
Flows and Dry Weather Flows resulted in completion in 19S7 of QtlLE, a 
computer model capable of reproducing water quantity transfer on a tile- 
drained field and in a small agricultural basin. This model, which was 
calibracad and verified on two tiled fields in southern Ontario, poBsesses 
physically -based parameters which may be easily determined independent of 
the modal itself. Excellent reproduction of flow peaks, volumes and 
hydrograph shapes was obtained with QTILE. 


A»s»ssiD«nt of Che Impaccs of clle drainage «y»c«B» on p«alt ■nd low 
flows ac Ch« flalti and cniall basin levels resulted In che conclusion chat 
tile drainage did not »tgnlf Icancly alter the cocal runoff voluae bviC did 
change the relaciva magnitudes of surface and subsurface flows. Because of 
this alteration in flow paths to receiving acreans. processes related to the 
presence of water, such as soil erosion, nutrient transport and herbicide 
and insecticide transport and decay aechanlsias. night' also be significantly 

Research waa continued in order to link quality algorithms to QTILE to 
assess the iiDpacts of tile drainage on water quality in receiving streams. 

After consideration of the state-of-the-art in quality simulation as 
It relates zo agricultural watersheds, the following research objectives 
wete defined: 

1) definition of the proceaaes Involved in novenenc of contaminants 
through the aoil or over the surface and into the dies or ditches 
draining agricultural fields, 

2) Incorporacion of an understanding of these processes into a physically- 
based model capable of simulating water quality changes on a basin 

3) collection of field data for calibration and verification of the model, 
4> use of the model to ovaluace the effects of tile drainage on sediment 

and water quality loadings, and 
5) provision of guidance on the use of the nodal for evaluation of 
pocential menageniant ECrstegle*. 

This poster presentation provides an opporrunlty to describe the 
progress made in achieving these objectives. 

A comprehensive review of existing literature was performed in order to 
summarize present knowledge of the processes controlling contaminant fates 
in agricultural systems. Contaminants of Interest fall into two groups: 
nutrients . a grouping which includes common agricultural forms of nitrogen 
and phosphorus, and pesticides, which includes a wide variety of herbicides 
and insecticides, Information was collected about sediment transport 


piroc esses, adsorpEion-desocption rcACtions . soluclon-precipltacion 
le actions , comp lexaclon . cnlccobiological caacclons, plant upt«ke, 
volati. Illation, ptiotclysls and hydrolysis for chei* two types of 
contanlnants . Thei a processes opera ta at different levels of Inportanc* 
depending on the specific contaislnant and physical systea under study. 

The premise of this resaarch is the u.e« of the developed quar^clty 
model as a framework for water qualicy submodels. Hence , preliminary 
modelling efforts have been directed towards a representation of the 
physical system which considers chemical trans fomat Ion of the concaminanc 
of interest as a sink term within a transport Bass balance provided by the 
QTILE quantity model. Later efforts will adapt these water quality 
algorithms for more sophisticated treatmenc of specific contaminants. 

During development of QTILE. a tiled research field wast of Kingston 
near Kapanee, Ontario was instrumented to provide data for model calibration 
and verification. A collection tank with compound V-notch welt at the tile 
outlet was installed to monitor tlla discharge. Rainfall volunes and rates 
were determined using a tipping bucket rain gauge and continuous strip chart 

To gather data for the development of water quality algorithms , this 
monitoring of the Kapanee field has been continued and extended. A 2i 
bottle water quality sampler (ISCO Model 2100) . modified to commence 
sampling once a threshold discharge from the tiled field has been exceeded, 
has been inscalled. Following the collection of samples, arrangements have 
been made for their analysis for contaminant concentrations. 

In addition to the Napanec field, a small basin (1.5 kn' ) In a corn 
growing area of the U'ilton Creek watershed, has been instrumented for 
rainfall, runoff and water quality sampling. 



OF ONTARIO RIVERS. Wm. C. Allison, T.E. Unny, University of 
Waterloo, Waterloo, Ontario, N2L 3G1. L. Logan, Environment 

The terra EXPERT SYSTEM has been applied indiscriminately to many 
diverse types of computer programs. It has become a "catch-all" 
phrase for any software that provides the user with more than a 
numerical response to a problem. The label EXPERT SYSTEM, has 
evolved into a cliche that is over-used and perhaps not well 
understood . 

"Machine Intelligence" distinguishes true Expert Systems from 
deterministic computer models. "Machine Intelligence" is the 
ability of a computer to reach complex inferential solutions to 
problems. The computer must be capable of searching through many 
heuristics {rules of thumb) and selecting the appropriate rules 
that suit each individual situation. A more suitable title for 
an Expert System which exhibits machine intelligence is an 
Intelligent Knowledge Based system (IKBS) . This name clearly 
implies that the system contains knowledge that emulates the 
information stored in the human brain. Searching through a large 
array of heuristics is the computer eguivalent to the human 
thought process. 

The analysis of water quality data is a complex task and it is 
often difficult for hydrologists to accurately and consistently 
interpret the results. WatQUAS 2.0 is an Intelligent Knowledge 
Based System for the assessment of water guality in Ontario 
Rivers, A comprehensive numerical analysis is conducted on the 
historical water quality record of a river monitoring site. An 
expert interpretation of the water qual ity at the site is 
completed by utilizing the results from the numerical analysis, a 
large knowledge base and an inferential engine. Conclusions 
regarding the origins, seriousness and possible solutions to the 
water pollution problems are presented by WatQUAS 2.0. 

Large quantities of heuristics and domain and expert knowledge 
are utilized by the Expert System to produce inferential 
conclusions and recommendations regarding the water qual ity at 
the monitoring site. WatQUAS 2.0 possesses a large knowledge 
base which contains detailed information pertaining to many 
conventional , organic, and bacteriological pollutants , 
Approximately 2 55 di f f erent pol lutants are contained in the 
knowledge base. The Canadian Water Quality Guidelines document 
was an important source of information relating to contaminants 
in the aquatic environment. 

The Ontario Ministry of the Environment "Effluent Monitoring 
Priority Pollutants List" lEMPPL) was utilized as the basis for 
determining which contaminants presented a danger to Ontario 
waters. The chemical hazard ratinq system developed for the 
EMPPL is utilized by WatQUAS 2.0 to determine the specific 
problems presented by each contaminant. 


A Data Base Management System is util ized to organize the 

knowledge block and to permit ease of access to its contents by 
the users. This feature allows for continual updating and 
expansion of the knowledge base by the domain expert. RAM 
requirements for the computer system are also minimized because 
the Expert System utilizes only the knowledge immediately 

The knowledge base and heuristics have been greatly expanded and 
reorganized in this second version of the Expert System. The 
similar format of the rules for each parameter was recognized in 
the construction of version 2.0 and rule frames were developed. 
For most water quality situations only one general rule frame 
with the specific information being retrieved from the Data Base 
Management System was used for all parameters 

A thorough statistical analysis is conducted by WatQUAS on the 

water quality data. Both parametric and non-parametric 

statistical techniques are employed to insure a complete and 
unbiased analysis. 

A new Water Quality Index was developed for WatQUAS 2.0. The new 
index examines and aggregates the various impacts that each 
pollutant detected at a site presents to the environment - 

WatQUAS 2.0 utilizes the Beale ratio estimator for the 
calculation of pollutant loads. This is the same method utilized 
by the Ontario Ministry of the Environment. This ratio estimator 
permits the Expert System to calculate loads which are more 
accurate than those calculated by WatQUAS 1.0 and are fully 
compatible with the requirements of MOE. 

The ratio estimator technique also permits WatQUAS 2,0 to 
identi fy pollution sources . The quantity of point-source 
pollution is calculated in the flow stratum representing the base 
flow. Once identified, the quantity of point-source pollution is 
subtracted from the total quantity of pollution in non-base flow 
strata to yield the total non-point source pollution load. 

Hypothetical pollutant load reduction is also examined by WatQUAS 
2 , Revised pollutant load estimates are calculated by 
utilizing a percentage reduction in pollution. This permits 
water quality managers to examine the effects of control measures 
and abatement strategies. 

WatQUAS 2.0 operates on an IBM compatible micro-computer and is 
intended for use throughout the Province of Ontario by the 
Ministry of the Environment. Some computer programming work 
remains to be completed that will link together the modules that 
compose WatQUAS 2.0. Future work on the Expert System entails 
enlargement of the knowledge base, increasing the number of 
heuristics and expansion of the scope of the WatQUAS system. 




ASSOCIATIONS IN THE DON RIVER. Lesley Warren* and A.P. Zim- 
merman, Dcpanmcnl of Zoology. University of Toionio, TDronio, 
Oniario MJS lAl. 

The total conccniration of trace metals found in ihc rceciving watets of 
the Metropolitan Toronto Area arc indicative of a icrious contamination 
problem. Il has become increasingly evident ihat assessment of ihe 
environmental impact of loadings depends more on knowledge of 
mcial spcciaiion rather than lowl concentration. Significant fractions of 
trace metals end up bound to suspended panicles with their ultimate fate 
{burial, rcsuspcnsion, tiioaccumulaiion vu:) connected to ttic fate of the 
system's particulate fraction. OMOE has articulated a need to determine 
the physical and ehcniieal eharacteristics of suspended particulate! in 
order lo assess accurately the itnpaci of metals on aquatic ecosystems. W'c 
arc in the process of evaluating A), the magnitude of metal transport by 
suspended sediments in the Don River; B). the gcochemical associations 
of metals with suspended sediments: C), the relationships between pani- 
cle size, meial load, and gcochemical phase; and D). if particle siie or 
gcochemical phase have any predictive power for flltcrfceding bcnihic 
body burdens of mcials. Four sites along the Don. progressing from ihc 
headwaters to the Bloor Street Viaduct arc under invcsiigaiion. 
Suspended sediments from these sites were coneentraied using continuous 
flow ccntrifugaiion and geochcmically characiCfiKd using sequential 
extraction. Pcciodically at one site, suspended material was size frac- 
tionated and gcochemical assaciaiions were analyzed within panicle size 
classes. The associations of copper, cadmium, zinc and lead with specific 
gcochemical classes were dciermmcd using Hamc atomic absorption. 
Body burdens of in-aitu niicr-fccding benthos were analysed for the same 
4 mcials. Early results indicate mcial levels in suspended material exceed 
provincial guidelines for dicdgeaic material (ranges across the 4 sites in 
ug/g: cadmium !,»>- 74; copper 18 - •ISO. line 4:o - 7O00: lead 60 - ^'^OO). 
or the 4 mcials. cadmium shows the itiosi predictable pailcrn of binding; 
it was only associated uiih the easily exchangeable and iron, manganese 
oxide fractions. Copper and lead show the most variable paiicrns of gco- 
chemical association with highest levels of copper appearing either in the 
organic, residual ot oxides Iraeiions. while lead levels were highest in ihc 
residual or oxides fractions 



ENVIRONMENTAL MONIIOftiNO C.A. Jefferson, Curry Jefferson Environmenlal 
Services, R.R * 4, Port Perry, Onlor to LOB INO 

The problem addressed by thisproiactwastrK availability of a single, x group 
of biomonilor ing organisms , which could be used in routtne cortam inant 
monitoring programs 

Proj-ams, requiring Oiscfwrge monitoring of coniamlnani Iwels in biological 
orgwiisms todetermtre impact trends wilh lime and space and pinpoint 
contaminant sources, ere expanding Ihe« programs require a common basis (or 
COfnptrison Conseoucntlydrgftni'm'. prwnn tn provide reliable, consistent and 
scientifically defensible data, and which are inenpensive tools, must be furlher 
dEvelDped and uIili.M 

The specific objectives of the pro)eci were (o 

-determine if one species oforganisfii wii rrLu:tipprjpr isle for biomonttoring, 
particulerly wilh respect toe specific dibchacge type; 

-to recommend one species of organism or a combination for either a specific 
siluMion.orgroupofconlaninants. to suggeit further research needs. 

The project involved discussions with researchers, a comprehensive and 
extensive review of the literature from inter naiional , provincial , and federal 
governments , industry, and educational institutions 


The bulk ol (He literature pertains to tr»e marine environment wUh particular 
ri^ca:! tomolluas The tresfiwaferdataemphtnL-e, irwmuHusc Wild the 
eii^eplion of work under lakwt &y tne Onlarm riinistry of ine Environment , no 
scltddat^jase of orwipffl-ticular species or particuls-ccnlamlnantfiasboon 
(tevelopea L llerature on the use of aitjse, Dentnic organisms otrter than Bivalves, 
^ixtpiankton. macrophytes, and fish Is not widely avaiiaie 

Ihe Ontario Mimstry of the Environment has the best datsUse of 

The project findings suflaest XfOt- 

-r;.vn<; np rfli»m«1;»r<T l«W!hw further rlftvplnfwrtw htnmf»iilnrifvinrOBrKm* for 
relatively stiwi-iei'm itudiesf I w<y1 i.\ i,r„-,nth^), 

-clamt De ulihiad for certain motrti',,'., fur w,, isrBi.'rt-i.3flniV,filfMino 
itirrlwilnmis. while leecfKS OedeveloptsJ fw L-tilwijphtiixjIi, 

procedures te developed for tne use of these two organismj, 

-Ihe small year iingveHnwoercn/SDOdaiUhiner; continue to be j=;ed 9^- 
loog-term ( -4 months to 1 1/2 years) Biomonitors of wider ge>V(»hic scope 

-ciams and leeches can be used singly or m tandem to pinpoint and define the 
nature and souri* of the specific contaminant problem. 




H. Vandermeulen and K.H. Nicholls 

Ontario Ministry of the Environment , Water Resources Branch, 

Box 213, Rexdale, Ontario, Canada H9W 5L1 

While Ontario's phosphorus control prograia has achieved 
measurable reductions In blue-green algal blooms in some inland 
lakes, there exist many other surface waters for which 
convent ional nutrient loading controls are not practical . One of 
the objectives of the inland lakes program is to apply other 
methods to control excessive growths of blue-green algae in 
eutrophic lakes and reservoirs in Ontario. We are using the 
"whole lake" approach to acconpl ish this task. Three different 
projects were initiated during the summer of 1988, 

destratif icat ion using a Garton style propeller with an upwelllng 
tube at Ouelph Lake, hypol imnetic aeration at a small kettle lake 
in London and a calcium carbonate addition to Puslinch Lake near 
Cambridge , 

Destratif ication - In late September 1988 a 6 m X 6 m raft of 
welded angle iron with a wire grid deck and ten 230 litre metal 
drums for floatation was anchored over one of the deepest 
sections of Guelph Lake. A 4 m long (3.5 m diameter) tube 
constructed of 2 mm thick polystyrene sheets was attached 
vertically to the raft 2 o from the lake surface. A metal 
framework reinforced by guy wires was used to suspend a propeller 
driven by a submersible electric motor inside the tube, 1 m from 
the top. The propeller ayBten (Flygt Canada, model 4410 Flomaker] 
utilized a 3.2 HP motor (600 Volt, three phase) with attached 
gearbo.\ producing a propeller speed of 32.3 RPM. The propeller 
was 2.2 B in diameter and used "Banana" style blades which were 
designed to produce a flow rate of 2.9 m^ / s. The motor was 


connected to an electrical supply within the Guelph Lake dam via 
a 250 tn long submersible SOW cable. The destratif ication system 
vas designed to move bottom water up to the surface of the lake 
for gas exchange. The system will be removed each winter for 
storage . 

Hypolianetic Aeration - In early October 19B8 a 6.1 m X 2.5 m 
raft of welded angle iron with a plywood deck and ten 230 litre 
metal drums for floatation was anchored over the deepest section 
of a kettlf lakff in London. A 5.5 n X 1 . 8 m X O.S m deep mixing 
box constructed of l.B mm polystyrene sheets was placed inside 
the raft and covered by styrofoam SM RiO insulation (10 cm thick 
on sides, 15 cm thick on bottom ) . Nine PVC pi pea (1.3 cm 
diameter, l.B m long, with 1 mm diameter holes drilled every 2.3 
cm ) were laid across the bottom of the mixing chamber at 0.5 m 
intervals. Baffles were placed between the pipes. The pipes were 
connected to a 0.75 HP oilless compressor (240 volt, three phase, 
!■ SCFM, 100 PSIl bolted to the deck of the raft. An B m long (38 
cm diameter! polystyrene inflow tube was attached vertically to 
one end of the mixing box. A 7 m long (46 cm diameter) 
I'olystyrene outflow tube was attached vertically to the other end 
.f the mixing box. The end of the outflow tube was angled to 
discharge water away from the raft. A metal framework was used to 
s'ispend a propeller driven by a submersible electric motor inside 
the inflow tube, 30 cm from the top. The propeller system (Flygt 
Canada, model 4400) utilized a ] . 5 HP motor (240 Volt, three 
phft^e I producing a propeller speed of 1130 RPM. The propeller wns 
2Z cm 111 diameter find was designed to produce a flow rate of 0.1 
m^ / s. The compressor and propeller motor were connected to a 
domestic electrical supply (Rotophase converter used to change 
single phflse to three phase) at the lake shore via a 200 m long 
=.ubmersibte SOW cnble. The propeller pulled water up from the 
hypolimnlon into the mixing box were air supplied via the pVC 
pipes agitated the water and baffles forced it along n 
circuitous path to enhance gas exchange. The oxygenated water 
was returned to the hypolimnion by gravity via the outfow pipe. 
The Bvstem will be removed each winter for storage. 


Caicium Carbonate Addition - In early Scptt^mber 1 98B , powdered 
iimoBtone [ 96X CaCOi , SOX of particles ' -t 5 urn 1 was brought to 
Puslinch Lake via tanker truck and blown into the hold of a 
apoe-ialij- designed trimaran vessel. The -vessel had a 
computerized pumping syalen to take on lake water and mix it with 
the powder in the hold . The result ing slurry wna deposited on the 
Burfnce of Ihf lake via spray arms rattnched to the stern of the 
sessel cohering a width of IH m. Moat areas of the lake deeper 
than 1 m were si-^rnycd in this mnnner . A total of 7B metric tonnps 
of I imp St. on t' powijer wo3 applied within three days. 

The- dostratifiiration of a portion of Gu'?Iph lake will continue 
^luring the s-imm'-r of 19ea. The testing of this aystem during 
September I9H8 indicated thut it does function but the size of 
the efferted area of the lake nec-ds to he detemined. It is hoped 
'hnl dPStrnl 1 f icnt ion viil reduce the severity of blue-green 
iilgdl bloDHs In th"? Inke ns has been reported by other authors. 
fieatratif Icat ion <~tin present to anoxic hypolianion from forming. 
Vnoxif: cotidit ioi'iB in bnttom waters during the Buminer months 
eiii^ou rages sed iment release of nutrients ond metal loiis, n 
|i|ieii(iD('non uhich is correlnt ed v ith Idri'. es*J the ^nsct of blue- 
gi-'-en Hooms . 

Tlie t-stins of the hypo 1 imn'-t ir mritioii ■system in 'he kettle 
lalic has n'lt been ecmpletcd at the time of writing. The system 
will be run durtn^ the ice free period of ISHS to .ietcrmine its 
effet-; upon the Inlif niid litue-grecn -nlgnc. Hypolimnetic aeration 
i 'i alio ONpei-trd to prevent sediment r^'lease of nutrients, it 
ill f f (.'rs from des t r'l*. i f icn t i on i ri t h«l ! he hyj^ol i nini en is 
pr'-^erved. The bottom unters -an then be naed far maintaining a 
t.ild Writer fishery in 'i lake were Jow oxygen levels previously 
pre\ented Hs eat nbl i sh«ent ■ 

The addition jf ( alt ium corhonate to F'uslinch LaUc should not 
have a measurable "ffect upon the lake until the svimmer of 1S8S. 
51ow dissolution r>f the t-nicium carbon.ite during the winter of 
19U8 will rrleasi- calclun ions whieh will bind with phosphorus 
nu<l lead t;i i\ I nw cnritrntrii ion rif Hi i ^. niii neni . l>;irm water 




TO MXiM}LJvric»s (f tciAPiirric t'lutfcmous ause. 

P.M. stokes, R.L. France and E.T. Howell. l.'^cifJte for Envirormental Studies 
and D^3t. of Botany, Uni-^ersity of Toronto, Toronto, Ontario MSS 1A4. 


One of the most visually con^icuous of tlie biological changes related to 
acidification of surface waters is t)ie development of accumalations of 
benthic filamentous green algae. These growths are canposed mainly of algae 
from the family Zygnemataceae, and principally frcm the genera Mpijqeotj, ^ and 
2iacsaailffl)' Frequently the growths are metafiiytic, appearing as loosely 
attached clouds ip to 2m in diameter, distributed son^irtTat heterogeneous ly 
through the littoral zone. The phenomemon has attracted the attention of the 
public, for the metaphytic gi-owths appear to have been increasing recently in 
small aoftwater recreational lakes on the Canadian Shield. In a 1986 survey 
conducted for the Csitario Ministry of the EnviroTBTient (CM)E) of more than 
5,000 cottagers in Che Parry Sound-Muskoka-Haliburton area of Ontario, the 
researchers concluded that approximately half of the study lakes had some 
metaf*iytic algal growth, and chat in 16% of the lakes, the growths were 
"considerable" . 

The phenanenOTi is quite widespread, having been identified in acidifying lakes 
and streams in southern Sweden, southern Nonray, the Mirondacks in New York 
state, and New Hairpshire lakes as well as in south -central Ontario. 
EKperiitiental acidification has frequently produced sijnilar although not 
identical effects. Although excessive develc^OTHit of the zygnesratacean 
metaphyton is not restricted to acidifying waters, t.'-ere Is neverUiless a 
certain consistency in its occurrence, and a nuiter of hypotteses have been 
put forward to explain this. One of these hypotheses, and the one which 
provided the major inpetus for the present stucfy, is that diininished grazing 
by herbivores results in less removal of the algae in acidic than in neutral 
waters, praaxing an increase in starKiLng bicmass. This says nothing ^asut 
the productivity of the cormiunity, whicii is the subject of another hypothesis, 
and of related studies in our gro^a. 

Several of t^le experiirental studies on acidifleaticgi tiave concluded or 
suggested chat scne (unmeasuied) decrease in benthic grazing pressure may have 
contributed to the observed develcprent of algal .tats. Certainly a nurter of 
zocbenthic organisms which have the potmtial to ccffisime filameitous algae are 
adversely affected bi' acidification, from pH 6.0 and below, but to date the 
si^xxsrt for the hypothesis has been inferential, 


The present study set out to: 

(a) characterize the algal bicmass and potential herbivore fauna in five 
Haliburton-Muskoka lakes, three of which were known to develop extensive 
zygnematacean clouds, and 

(b) forward atn explanation for the algal accumulation and persistence in 
relation to the abundance of micro- meio- and macro-f au-nal grazers . 



In the sumier of 1997, the five sti*±/ lakes were selected on the basis of a 
priori knowlec^ of Che presence and absence of filamentous green algal 
clouds. The lakes have been variously described and studied by the CWDE, and 
will not be described in detail here. They are: Plastic Lake {fH 5.6, has 
met^ytic clouds = PL), Gullfeather Lake (pH 5.9, has metaphytic clouds = 
O"), Bentshoe Lake ipti 5.9, has TOtapiiytic clouds = BS), Crosson Lake tp« 5.7 
- 5.0, has so far not develcped rataphytic clouds - CL) and Little Clear Lake 
(jaj 6.5, no mete^ytic clouds = LC) . 

Lakes were sanpled over the period June - August 1987, with each lake being 
sanpled on two occasions. Met^ihyton was sanpled quantitatively and the three 
types of grazers, grouped for conver.ience as "micro-" (cladccerons, 
oligochaetes, chironomids etc.) , "meio-" (atnphtpods, gastropods, 
ephemerc^terans etc . ) and "rracro-" {crayfish, tadpoles and small minnows) 
were sanpled by six different techniques in total. Substrate was taken into 
account, with major macrojdiyte canmunities ar,d sediment type being recorded. 


1. MetacJiyttTTi biemaas . A sunmary of the bicmass of mecaphyton is given in 
Table 1. There was extensive growth of metapiiyton in Plastic and Bentshoe 
Lakes, dominated by Zygogonium tunetatum . Gullfeather had less growth, and 
the clouds were restricted to specific locations, while no such clouds were 
found in Crosson and Little Clear Lakes. The time of appearance of the 
iret^Siyton varied fron lake to lake (Table 1) . 

Table 1. Bicmass of metafiiytic filamentous green algae in the 0-2 m d^xh 
interval in the study lakes. (N=80) 

Lake Hame^ 





aiomaSB of metc^ytcm 
g m~2 d.w. (std. errl 


0.83 (0.20) 



0.97 i0.20) 

1.21 (0.29) 

shoreline however this ctensity of material was considered to low to 

^See text for full names of lakes. 


2. Grazer abiirrtanf-pq . 'mere were no overt differ&ices in the aixindances of 
micro- cr raeio-grazers amor.g the lakes. Notable differences did exist 
however, witli respect to the macrograzers . Crayfish vlrilis i were 
ccmon in Little Clear Lake, abundant in Crosson Lake (Cantoanjs barton! . Q_^ 
viriliq l , but absent or rare ir. the three lakes characterized by retaf^ytic 
algae. Tailzies (pana cl^mitans i were also abundant in the non-nietaphyt<Bi 
lakes - Little Clear and Crosson, while absent from Gullfeather and Bentshoe 
Lakes vrfiich had Z ygocionium . Tadpole data for Plastic Lake are inconsistent 
with the macro-grazer - mecaphyton complementarity. Lakes without metaphyton 
tended to contain greater relative abundances of algivorous fish than those 
lakes in which algal developnents are present. Table 2 sunmarizes the data 
for macrograzers . The most notable is Crosson, with a low pH, but with 
abundant crayfish and no metaphytic clouds. 

Table 2. Abundance of macro -grazers in Haliburton^*JSkoka lakes. Values in 
parehth.e3as denote the average standard error of SCJBA determined densities or 
trap catches between the two census periods. An enprical estimate of crayfish 
abundance was derived from trap catches with the equationof Capelli (1975) as 
used by France (1965). Data for fis^i represe."t ordinal rankings calculated 
for cyprinids. 

Crayfish Tad^^les Fish 

Lake No, m"^ No./trapi^ Enp.No. m~^ No. m ^ No. /trap Traps 





























1 = number of adult males 


If alterations in grazing pressure can be inplicated as an inprotant attribute 
underlying metaphyton develcpmer,t, our correlative results suggest that it is 
these algae grazers (particularly crayfish) that will be the rrost likely 
candidates. A considerable literature exists providing inferential support 
for this hypothesis. During 1988, we have surveyed a total of 38 lakes to 
determine the strength of the relationship observed in IQSl. 


The authors ackiiowledge financial and logistic SL^port from the CM3E (Research 
Advisory Cortnittee, Dorset Research Centre) and appreciate scientific 
discussion with Michael Jackson, Michael Turner and Norman Van. 




Hariusz Rybak, Izabela Rybak 

180 Elgin St., Ottawa K2P 2K3 


Kenneth Klcholls 

Ontario Ministry of the Environment 

Water Resources Branch 

125 Resources Road 

Rexdale, Ontario 

Presented at the 1988 Technology Transfer Conference 

Toronto, Ontario 

November 1988 

* The full text of the Report was submitted for publication to 



Relationship between surface sediment cyst assemblages and 
laka-water characteristics were studied in 50 lakes located in 
central Ontario. The main purposes of the study were to identify 
the environmental factors most strongly controlling the distribu- 
tion of chrysophycean cysts and to develop indices and equations 
to infer lake water pH from cyst assemblages. 

Surface sediment samples were collected from a total of 50 
lakes in the Killarney-Perry Sound-Muskoka-Hallburton region of 
Ontario. The lakes were chosen to represent a wide variety of 
central Ontario lake-waterahed syatems (see also Griffiths e£ 
al., 1988) 

Principal components analysis indicates that TDS and as- 
sociated lakewater pH as well as elements related to trophic 
status are the most important factors controlling the distribu- 
tion of chrysophycean cysts. There are significant differences in 
the relative importance of these factors among the lakes. 
Generally, the large number of significant correlations suggests 
that chrysophycean cyst distribution is influenced by one or more 
of the six environmental factors. Most of the morphotypes seem to 
be significantly correlated with alkalinity and lakewater pH (21 
taxa) , while 19 morphotypes appear to be correlated with trophic 
status of the lake. The results suggests that due to complexity 
of variables such as pH and trophic status, which can be in- 
fluenced by several environmental interactions, there could be 
differences in the level of importance of several environmental 
factors controlling the distribution of cysts. 

Several techniques were used to develop equations for infer- 
ring lakewater pH from fossil chrysophycean cyst assemblages. 
Calibration equations l and 2 (Table 1) predicted surface-water 
pH most adequately and the precision of those equations was the 


l*le 1 »«reMiwi equation tof iofefrinj {M tnm fMiil ehrwfltfW""" ty»t •»B*"til«9»»- 
«n raldMnshlp art hlgniy siv^i'icmi (l-<D.D001) 


1. p«.7.07'0.19JC,^-0.1SJCj^-0.(BlCj2-O.Mn^'0.ir6Cg3-Ci.H«^-0.10«^-0.280C,o^-0.3Jl^^ 

2. ^•l.BS6-D.1Ilk^0.02Ita:fD.01UI<'a.OS1UM4.029MIU> 

WW* C^ - X abinliifre of IM fn) ■t>rtf>o(VDa 

tall - X abindinc* of icidohionttt lorm 
Mf - X tbunOtrct at »cnlDf*>iiixrt form 
t • X abinlwic* of IndifterenI furaa 
«lkf' X aburdanc* of Alhalif^tiiloui form 
«llib- % BtxnUrct of HUalibionllc form 

The equations can be recommended for paleoecological studies and 
are applicable for investigations of a wide pH range of Ontario 
lake types. It should be noticed that in order to increase the 
confidence of ecological interpretations based on the derived 
equations, it was necessary to estimate the relative precisions 
of cyst-predicted values using data from lakes that were not a 
part of the calibration lake set (.i.e. the validations lakes); 
Raven, Maggie, Papineau, Diamond and White). The regression of 
the pH predicted from equation 1 against the measured has a very 
high correlation coefficient (R=0.99); SE+).18) and the pH es- 
timated by this equation does not differ significantly (i.e. the 
intercept and the slopes do differ significantly from zero and 
one, respectively) from that measured in the validation lakes. 
Although the relationship between the predicted and measured pH 
in the validation lakes have a very high R values based on other 
derived equations. 


The study also provides a descriptive analysis of the 
"fossil" chrysophycean cyst flora from Ontario lakes. The 
descriptions include representative SEM micrographs and detailed 
characterization of each morphotype in consideration of the mor- 
phologlcal variation observed among specimens of the same mor- 
photype. special attention has been paid to the anatomy of the 
collar complex and to the nature of the cyst surface ornamenta- 
tion. One hundred thirty seven inorphotypes are described, most 
of them for the first time. 

The present study as well as previous investigations (Rybak, 
1986; Rybak, 1987; Rybak et al - . 1987) show a great potential for 
using chrysophycean cysts as paleoindicators. 


Griffiths, R.W., Carney, E. , Nakamoto, L. & K.H. Nicholls, 1998. 
Parametrization of calibration equations relating diatom 
"microfossil " to the surface-water pH and alkalinity in 
central Ontario lakes. Ecol . Honogr. 

Rybak, M., 1986. The chrysophycean paleocyst flora of the bottom 
sediments of Kortowskie Lakes, Poland and their ecological 
significance. Hydrobiologia 140: 67-84. 

Rybak, n. , 1987. Fossil chrysophycean cyst flora of Racze Lake, 
Wolin Island (Poland) in relation to paleoenvironraental con- 
ditions. Hydrobiologia 150: 257-272. 

Rybak, M., Rybak, I. and M. Dickman, 1937. Fossil chrysophycean 
cyst flora in a small meromictic lake in southern Ontario 
and its paleoecologlcal interpretations. Can. J. Bot. 65: 



Hebert , Craig. E. and G .O.Haf f ner . 1988 . Ecological partitioning of 
organochlorinated contaminants in forage fish species. 
Great Lakes Institute, University of Windsor, Windsor, 
Ontario. N9B 3P4 

Three species of forage fish : l-abidesthes sicculus (Brook 

Silverside). H fftropis hudsonius {Spottail Shiner), and Pimephales 

pQtatus (Bluntnose Minnow) were collected from three sites along 

the St. Clafr fiiver systeir during July, August, and September 

1987. The fish were caught in nearshore waters using a 0.6 cm 

tnesh bagseine. They were measured (total length) and immediately 

wrapped in hexane-rinsed aluminum foil. Samp les were kept frozen 

at -20 degrees Celsius until they were analyzed. 

The three species are morphologically distinct and it is 
these physical differences that separate them ecologically (Keast 
1966). L- sicculus is primarily a surface feeding species whose 
beak-like snout and dorso-lerminal mouth are designed to seize 
prey at the surface. P. notatus possesses a ventro-termi nal 
mouth designed for benthic feeding. N. h udsonius has a terminal 
mouth and is more omnivorous than the other two species. 
Therefore, there was an integration of the foodweb with L- 
sicculus tracking a 1 1 ocht honous surface inputs, H. hudsonili^ 
pelagic food resources, and P. notatus benthic foodwebs. There 
would have been some overlap in resource utilization but the 
special ized feeding adaptations of L. sicculus and £. not.a^Mi. 
would hasie limited this resource sharing. 

Whole fish were analyzed according to the protocol developed 
by the Canadian Wildlife Service (1982). The samples were 
injected into a GC-ECD. Pentachlorobenzene (log Ko„ = 5.0) , 


factor regulating contaminant levels in forage fish. This was 
especially true for the higher Kq^ compounds, HCB and OCS. 
Bioconcentration, however , may have been more important for the 
lower Kq^ compound, QCB. This would have accounted for the 
smaller interspecific difference that was seen for this compound 
in that QCB exposure would have been more homogeneous among the 
three species. Due to their relatively high Kq^ and low aqueous 
solubi I ity HCB and particularly OCS would be expected to 
partition more readily into sediment. When feeding, P. notatus is 
in continual contact with the substrate and may ingest a 
substantial amount of sediment with its food items. This was 
perhaps the best explanation as to why this species had 
consistently higher contaminant burdens particularly with respect 
to the higher K^^ compounds . It. h^Jsonius . a facultative 
omni vore. general ly had higher contaminant burdens than L.. 
^icculus but lower levels than £.. nptatuS - This might have been 
due to its selection of a wide variety of food items which varied 
in their degree of contamination. The levels of contaminants in 
L- sicculus remained much lower than in the other two species 
because a large proportion of its diet consisted of terrestrial 
insects with lower HCB and OCS levels. 

Ratios of contaminants are useful in that they yield 
information on foodweb interactions. Some organ ochlorine 
contaminants with high K^^s wilt partition more readily to 
benthic systems whereas others with greater aqueous solubi 1 ity 
and lower K^^ will remain in the pelagic system. The proportion 
of one type of compound to another will give an indication of 


where an organism is primarily feeding. Using this assumption, 
organochlorine compounds may be thought of as ecological markers 
highlighting foodweb interactions (Flint 1988). For L- S. U . Cf 1n? 
it was observed that the ratio of lower to higher Kqw compounds 
».as greater than the ratios observed for the other two species 
(Ratio HCB:OCS; L- notatus °0.4. H. MAkOAluX^ ^ ■ '^ • ^■ 
sicculus ^l .9) . If habitat utilization and therefore food 
selection was important to contaminant uptake then this would be 
expected. U- s i ecu! us was not exposed to OCS contaminated 
sediment or to foodwebs associated with benthic habitats due to 
its surface feeding behaviour. 

The interspecific differences that were observed indicated 
that habitat parti tioning was a major factor regulating 
contaminant levels in these forage fish species. Although 
chemical and physiological parameters may determine which 
contaminants have the potential to bioaccumul ate. it is the 
regulation of exposure through ecological processes that will 
determine the degree to which that potential is realized. 

pi bl iograohv 

Canadian Wildlife Serv ice . 1982 . Analysis of organochlor ine 

hydrocarbons in tissue samples. Analytical Manual. 

Environment Canada. Ottawa. Ont. _ 

Flint R. H..W.H. McDowell , and G . Yogi s . 1988. Potential use of 

organochlorine contaminants to validate a food web model. 

Verh. Internal. Verein.Limnol. 23:265-270. 
tteast A. and D. Webb . 1966 . Mouth and body form relative to feeding 

ecology in the fish fauna of a small lake, Lake Opinicon, 

Ontario. J . Fi sh . Res . Bd . Canada 23:1845-1874. 
Oliver 6. G. 1987. Bio-uptake of chlorinated hydrocarbons from 

laboratory-spiked and field sediments by oligochaete worms. 

Environ. Sci.Technol. 21:785-790. 




D.R. Van stempvoort, P. Fritz, and E.J. Reardon 

D«pt. of Earth Sciences, University oE Waterloo 
Waterloo. Ontario N2L IMS 

The environmental impact o£ acid rain sulphate has become an 
Issue ot both national and intifrnatlonal concern. The 

Plastic Lake watershed (Dorset, Ontario) is an acid raln- 
affccted site that has been th* subject of an intense 
geocheinical investigation by Envtronaent Ontario over the 
pair 8 year?-. The svilphate balance m upland forest soil at 
this &it«r can be .nodelled simply as a input cE sulphate from 
the atinofcpiiere, aiid output >jI sulphate to cji'otnidwater , 
neglecting ■jeochfemtcal piceA^^ii and transf ot matioiis wituin 
tlie soil. However, this broad ap(>roach is subjei^t to large 
•?i'iv.irs, .iurl. 3S lirge uni:ertainty in the tate of dry 
deposition of aimospheric SO; and paiticulare sulphai •:' 
S.Bliar.y, aOiE-r throiiyhf 3ll sulphate may havr- ;ear,h»^d Irim 
leaves ot plsnts r nat .iirorpoi «te-d r he .suliihrtr.* hy root s 
tiom poil,. Tliib pooi-ihility .■^re^te*. more uhcerto;nTy m tl.e 
-alrTulation c( Input atmospherl &iilp-iate. 

"ut th-rrm a'e, o .iniul ei of previous biogeochemical studief. 
i.ndl ate that soil silpha'^e is not "con =«rvatlvi?" , but is 
ii".c-.a'po;"ate"l ty p. suf ^ and miL robes, aud altc^ is a piodur-t 
:t inii^ribial aimers ' nation of :>ji1 orgariic conpounds. Some' 
t the i^oil wiite ulpUate may be adsorbed and retained 

1 <']i>[iiiit«lv by Ic HI and aluniLnun oxyhydvoxides . It remains 


unclear jnet how these procefises affect the impact of 

sulphate on f'^rest ecosystems. 

In this investigation, the dettUled dynamics of sulphate m 
th& upland forest ecosysteai at Plastic Lake (1936-8S) h.3Vfe 
b-jen documented, in part, by stable isotope analyses of both 
sulpliuf and oxygen In sulphate-. These isotope ratios 
provide direct tracers of the fate of atmospheric sulphate 
tn Che doll. In this ecosystem, there is very little 
variation in tli& average sulphur i-sotope ration (^^''s o/ur. 
iCDTlI r^t sulphate i,n vari..>us resei'volrs [rainfall '■l.S. 
throughf all, -4.1, stemflow. E bor.zon leachate, ♦■J.5), or 
fractlsuB oE soil oigatil'; sulphur (leave:^, +3.9; lltt-rr. 
1-5."; humus, *3.$;. This lack of variation refle'":ts the 
d'iMiiiaiice of atin.-jsphe;.r-de. ivi-d sulph it lu this eccsy&cem, 
..lid the n'jhiniportdhce of t^nlphiir lactope fractionating 
pi.jcesse.- . 

" irthermore. isotope neasuretnents indii:*te that mi. st CO-lOf 
■:.) of th^ fculphate ih th;.-oucjhfaii Is derived frow th*- 
atinospheie: Thio.iyliisll salph^tte (dvg. f^^C = ♦10'^ (SMOWl! stfmf 1j.w sulphate (avg.E'-^O *^.3 have bligh' ly 
liyhter o:<yger. than the precipitation liulphate lavg. t^^g = 
♦ 11,4 ), Since thr'iughfall maKejj up around 90-95 'i of 
upland forest soil tiif iltratl^.n, it is clear that leaching 
of soil-derived sulphate from abovegi'iund vegetation i» not 
a majcr conipc itenV. of the sulphur rycle. 


Of gieat slgnif n-aiicp, dissolved aiiJ watei" soluble sulphate 
111 the upland forest soil lias a distinct oxygen isotope 
composition lavg. 6^^0 = +5.5) lelatlv© to local 
precipitation sulphate (S^Bo = +11.4 ). This miist be due to 
chernicai •.•r biological reactions xu thi? soil. Adsoiption 
o£ sulphati? by aluminum and lion oxyliydioxides is a very 
important process in many acidic soils, includlncj the B 
horizon (sandy till| of the upland Podzol.s in tlie Plastic 
Lal:e watershed. Ikiwever, field data and our ongoing lab 
studies indicate that this process has a minor effect on the 
isotope composition of snlpiiate in soils. In snme of onr 
tests , dissolved sulphate is enriclied m 1^0 relative to 
sulphate adsorbed by iron oxyUydroxides, opposite in 
direction to that expev:ted lE adsorption was responsible for 
the soil sulpliate oxygen shift (dis&olved sulphate depleted 
in ISq), In £aL't, tilt- soil sulphate oxygen isotope shift is 
alif-ady de-tecteible in leachate samples from the uppermost 
litter -liUDiuE zone (avg. 61^0 = t5.8), wliere sulpliate 
adsorption is minimal . Clearly this indi-raCes that soil 
adsorption is not tlie cause of the observed oxygen slillt. 

Ill light of the above evidence, pianl and microbe mediated 
processes, Eocnssed in the llttei-humus zone, are likely 
r-?3ponsilile fia" th-^ soil sulphate oxygen isotope shift. 
Plant and microbe bioaESiniilation of sulphate is a 
ubiquitous process in soils. Reduced organic sulphur 





Hariusz Rybak, Izabela Rybak 


180 Elgin St., Ottawa, Canada K2P 2K3 

Presented at the 1988 Technology Transfer Conference 

Toronto, Ontario 

November 1988 


Lake Huskoka, the largest lake in the district of Muskoka, 
provides a representative example of lake ecosystem under 
anthropogenic stress. The lake is located in an acid sensitive 
region with limited neutralizing capacity. It is exposed to acid 
precipitation as well as acidic run-off water from the 
surrounding areas. The stability of the ecosystem is also 
effected by eutrophication and contamination processes. The lake 
has an area of localized nutrient enrichment which causes 
periodic algal blooms {Gravenhurst Bay) with all the negative 
environmental changes related to it. 

The primary objective of the study was to analyze and 
document long-term water quality changes in the lake with 
emphasis on heavy metal contamination and eutrophication 

The approach to this research was based on the application 
of several pa leoeco logical techniques. Detailed analysis of 
metals deposited In lake sediments was used to estimate the 
extent of lake contamination induced by erosional inputs and 
atmospheric deposition. Analysis of major oxides, supported by 
fossil pigment analysis provided an assessment of the degree of 
lake eutrophication as well as exchange of cations between the 
sediment and the water column (induced by changes in hypolimnetic 


oxygen regime). Fossil pigment analyses (as primary) was applied 
to reconstruct the history of the lake's productivity and changes 
in the past and present trophic status. The history of blue-green 
algal development in relation to the eutrophication process was 
documented using detailed quantitative fossil pigments analysis 
(myxoxanthophyll and oscillaxanthin) . Recent events were 
accurately dated using the Lead-210 radionuclide method. 


Trace metal contamination 

In this study, 88 (eighty eight) sediment samples were 
analyzed for the trace metals Co, Cd, Ni, Mn, Cu, Zn, Zr, Hg, and 
Pb. The data indicate a significant spatial variation of heavy 
metal contamination among 24 studied stations. The highest 
concentration of metals was recorded in the core sediment samples 
originating from Cooper Point. Many of the measurments exceeded 
the Ontario HOE guidelines. Most of the analyzed elements in 
sample core samples studied elements showed a systematic change 
in concentration with depth suggesting, anthropoganiclly related 
trace metal accumulation. 

Trophic status 

Stratigraphic shifts in the concentration of chlorophyll and 
carotenoids have often been interpreted as evidence of changes in 
primary production and trophic status. Host of the study sites in 
the lake showed similar levels of chlorophyll derivatives, total 
carotenoids and blue-green algal pigment concentration with the 
exception of Gravenhurst and Mus)toka Bays. In both cores from 
those two locations, the concentration of pigments showed 
successive increases upwards to the sediment surface. Only the 
most upper strata indicated a lower level of pigment 
concentration. This may suggest significant changes in the 
trophic status and level of primary production resulting from the 
phosphorus removal operation initiated in 1971 at the town of 



Metal contBfflinationof WetlandFoodchains in the Bay 
of Qulnte, Ontario A, Crowder", W, Dushenko and J. 

Greig, Dept. of Biology, Queen' sUniversity, 
Kingston, Ontario, 

In September, 1987, the Lake Situcoe Reqional and Metropolitan 
Toronto and Regional Conservat ion Authorities began research on 
the survival characteristics of fecal and water quality indicator 
bacteria in rural watersheds. Six locations within the confines 
o£ the LSRCA and MTRCA Rural Beaches Study areas were selected 
for examination. These sampling site locations included; 5 
sites in oeEferlnw rresk (LSRCM; 1 n i te in the Fflst Number 
River ; and 2 sites in Cent rev i lie Creek (MTRCA) , The bacterial 
parameters studied during the first year of this 2 1/2 year 
project included: pure cultures of Escherichia coli and 
pseudomonas aer^jginosa , and Cecal col if orms and Pseudomonas 
aecug i nosa Tn n i yed clj Itures containing total heterotrophic 
bacteria . 

The water column sqr'/ival of these bacteria has been assessed at 
each of the 6 sitf?s under 4 seasonal conditions: fall 
(September/October); winter (January/February); spring 
(April/May); and Rummer (.June/July). In addition to the water 
column work , sediment survival runs of E . co I i were completed at 
each of the sites under the above same seasonal conditions, 
making a total of IHO runs to date. Water column temperature and 
chemical parameters such as dissolved carbon; total phosphorous 
and nitrogen; nitrates; and ammonia were monitored during the 
runs to determine their effect on bacterial survival. Bed 
sediment nutrient levels and particle si zes were also analyzed. 
Invitro (Uaboratoryl survival exp-^riments were conducted to 
measure the effect of specific parameters, i.e. water temperature 
and nutrient content on bacterial survival under controlled 

Water column bacterial die-off rates in tho 3 watersheds ranged 
[rom P.l to 0,35 Log rjni ts/lay for F . Col i ; 9 .92 to C4 Log 
Un 1 1 s/Day for f aca 1 colitorms; 9.3 3-0.29 Log Uni ts/day for P. 
aeruginosa ; and n.iJl to B.25 Log '.'ni ts/day for P. aeruginosa 
(Mixed culturel. The rates exhibited were considerably slower 
than those reported for urban rivers and streams {fl.5 to l.P Log 
t]nits/day! (Seyfried, Harris and Young; 198fl unpublished). 

Some seasonal effect on the F . col i and fecal coli form rates was 
obser^'ed with more raoid die-off occurring during the summer 
(Pef ferlaw Creek) and spring (East Humber wiver and Centrevi He 
Ccpek) . Posults of the in- laboratory experiments conf i rm that 
more rapid die-off of EC and EC occurs under warmer temporal 
conditions. Rates for P. aeruginosa and P. aeruginosa (mixed 
culture) tended to be ■slower during warmer weather in Pefferlaw 
Creek however , this tr-?nd was not as evident in the 2 mtbCA 


E. coli die-off in laed sediments was less rapid than in the water 
column (0.01 to 0.1 Log Units/day) . Somewhat faster rates were 
nated during the winter in Pefferlaw Creek and during the spring 
in the East Number River and in Centrevil le Creek, Apparent 
regrowth of K. coli in the sediments was observed during the fall 
and summer survival runs in all 3 watersheds. This phenomena has 
been observed by previous investigators and is thought to result 
from the use of autoclaved sediments in conjunction with the 
warmer seasonal temperatures. 

Statistical comparisons of the die-off rates between si tes 
(d'Jrin-j -sn individual survival run) were performed by regression 
analysis. The results of these analyses show that bacterial 
survival in both the water column and bed sediments can vary 
within a watershed, and is thus site specific. 

The in-si tu relationship between water temperature, nutrient 
concentration and bacterial survival has not been fully 
established at this time. It would appe^ir though, that within 
the P.C/fC group, survival is somewhat enhanced when both water 
temperntute and nutrient levels ate in posi tive correlation i.e. 
high tempecature/hiqh nutrients or low temperature/low nutrients. 
This game trend was .si so exhibited during the invitro survival 
experiments. In- laboratory experiments to assess the effect of 
temperature and nutrient levels on P. aeruqimosa survival have 
iiot been completed yet . 


The methodology for conducting survival runs incorporated the use 
3f nembrane diffusion chambers developed by 0. McFetecs of the 
Montana state University. (McFeters and Stuart, 1972; 1981). 
Further modifications to the current method, including the design 
of the sediment and water column chamber holders, were made by G. 
T>3lmateer of the Ministry of the Fn/ironment, London, Ontario. 

Chambers for water column runs i/i?re prepared by inoculating 
bacterial cultures, diluted 10 fold in gterile ■site water, into 
sterile diffusion chambers. Time zero control samples weco taken 
and analyzed to determine the initial bactet ia 1 concent rat ion , 
The chambers were Chen transported in containers of chilled iA°C) 
site water to the test locations. Triplicate chambers of the 
bacterial suspensions were placed into anchored holders at each 

Sampling of the chambers was conducted over a two week period 
commencing with daily sampling during the first wee's of the run 

and 2 samplings during week 2. 


Samples were t ca n 3 poc ted to the Laboratory, on ice, where they 
were analyzed within 24 hours by membrane filtration (MF) using 
the following selective media; M-TEC for the determination of 
fecal conforms and E. coli (Dufout, 1981), M-PA foe the 
determination of P. aerunmosa (Standard Methods, 198 5) and MS PC 
I for the detetmi na t ion 57 total heterotrophic bacteria. 
(Standard Methods, 1985). 

Sediment survival chambers were prepared by inoculating 5 mis of 
an E . CO I i culture, diluted 10 fold in sterile site water, into 
chambers containing S0 grams of sterilized site sediment. Nine 
chambers were preoated pet .site, of these, 8 were transported to 
the field in chilled site water and the remaining chamber 
anal yzed as a time zero control. 

Sampl ing of the sediment chambers was accompl Ished by removing 1 
chamber from each site for analysis at 2A and 72 hours and once 
per week thereafter for 6 weeks. Sediment chamber samples were 
analyzed by MF or MPN technique depending on the bacteria level. 
FC «roth (Difico) + Methyl 'Jmbel 1 i f eryl-B-D-Clucoronide (MUG) 
(Peng and Hflrtman, 1992) was as the enrichment medium used in the 
MPN procedure. 

Invitro survival experiments were conducted using 10 gallon 
aquariums filled with water from Pefferlaw Creek. The tanks were 
placed in l«oc and 20°C incubators to achieve and maintain the 2 
desired temperatures. The preparation methods for water column 
survival chambers of E. coli and P. aeruginosa were identical to 
those used for the inside survival runs. Triplicate chambers of 
the bacteria were prepared for testing at both 10»C and 20Oc and 
were emersed in the tanks immediately. Sa-nplinq of the invitro 
chambers was conducted over a 7~day period and the samples were 
analyzed in the same manner as were the inside samples. 

Calculation of the bacterial die-off rates for both inside and 
invitro survival runs was performed by regression analysis of 
bacterial concentration with time. 



Oevelopnent of an Acute and Chronic Sedtaent Btoassay Protocol Using 
Larval Mayflies and Juvenile Fathead Minnows. Gail Krantiberg* and 
Richard Pope', 'Water Resources Branch, Ontario Ministry of the 
Envlronaent, Toronto, Ontario M4V IPS, *Tarandus Associates Inc.. 
21 Greystone Crescent, Braapton. Ontario L6G 2B2. 


Sediment bioassays measure the effects of contaminated sediments on the 
biota. Sediment elutriates have been prepared as liquid phase matrices, 
princlply to assess the impacts of dredging activities on water coluran 
organisms (1.2). For example, one toxicity test exposes Daphnia to an 
elutriate (3). Pore waters have been considered as an alternate liquid 
phase to examine the effects of contaminated sediments on the burrowing 
infauna and to Identify the route of exposure of different organisms to 
different pollutants (4). 

8y far the most frequently described approach is solid phase testing 
with either benthic or water column organiSBS (5,6). For the purpose of 
evaluating the impacts of Inplace pollutants on the biota, as opposed to 
the consequences arising from dredging operations, the focus of this 
study was on the solid phase bioassay. 

The principle objective of this study was to contribute to the 
development of a methodology for assessing the chronic and acute 
toxicity of sediments to biota. This included an examination of the 
effects of bioassay assembly and sediment manipulation techniques to the 
response of the test organisms, and the sensitivity of growth as a 
chronic endpoint. 

Experiment 1: To determine the efforts of settling time, following the 
addition of sediments and water to the bioassay container, on toxicity 
to mayf 1 ies. 

It is reasonable to expect that the exposure of an organism to 
contaminants will vary with the state to which the sediment-water system 
is in equilibrium. We therefore examined whether an organism's response 

varied with the length of settling lime of the bioassay assembly 


proceeding the Introduction of the organism. The duration of exposure 
required for the response of organisms in test sediments to differ 
significantly from the controls was also not known. As a result, the 
experiment was designed so that half of the replicates could Oe 
harvested at day 10 and half could be harvested at day 21. 

2L widemouth glass jars were filled to a depth of 3 era with sediment 
(surface area = 100 cm') and water was gently added. Organisms we^-e 
introduced at 3 time intervals; 5 hours settling plus I hour of 
aeration. 1 day settling plus 1 hour and 5 days settling plus 1 hour 
aeration. At each time Interval, eUher 8 mayflies {c.a. Zb mg/ 
Individual wet weight) or c.a. 1,5 gm oligochaetes wet weight (c.a, 150 
individuals) were added to the chambers. Each treatment had 4 
replicates. Water and sediment samples were collected as animals were 
added and when replicate containers were harvested (time = lOd or 21d). 

Analysis of the growth response of Hexagenia suggested that biomass 
changes were influenced both by sediment type and by the duration of the 
period of equilibration (Table 1). Eirowth in both test sediments was 
greatest when the mayflies were added 5 days after chamber assembly, 
followed by a I day equilibration period. Growth was poorest when 
organisms were added 6 hours following assembly (1 hour after aeration). 
Growth inhibition more pronounced by day 21. as compared to day 13. 

Experiment Z: To determine the effects of settling time on toxicity to 
fathead minnows at 2 different densities. 

Fathead minnows weighing c.a. O.S gm per inflividual were added to each 
bioassay chamber at a rate of 10 or 15 individuals per replicate. Four 
replicates of each treatment were harvested after 10 or 21 days 

In accordance with the biomass changes noted for mayflies, growth 
inhibition was least when the fathead minnows were added 5 days after 
chamber assembly. There appeared to be no notable differer.ce between 
the 6 hr. {5 hr. settling plus 1 hr, aeration) and 1 day equilibration 
periods with respect to biomass changes, and the effects of fish density 
were variable. Growth inhibition was greater with 15 as compared to 10 
fish in some. But not all cases, and density apparently exei-ted no 


influence on biomass changes in the controls. This last finding Is of 
interest, since it may indicate that the stress of possible overcrowding 
was exacerbated by the contaminated sediments. By day 21, all fish had 
decreased in weight. Minnows from the test sediments lost more weight 
than did the controls. 

E);periment 3: To compare the toxicity of intact sediment cores to 
homogenized sediments for mayfly nymphs and fathead minnows. 

CuTent methods for asierably of sediment bioassays often Involve sieving 
and homogeniiing the sediment. This effectively exposes the organisms 
to a unifom dose of contaminants that is in reality a mean dose of the 
heterogeniously distributed contaminants. In some cases, the extensive 
aeration of the sediment also results in a transformation of chemical 
species to forms that are of greater or lesser bioavailability. We 
examined the question of sediment homogeniiatton by using diver- 
col lected cores. The cores used were acrylic tubes of comparable 
surface area to the 2L glass jars. Organisms were introduced into the 
cores and into homogenized sediments from the same site as those where 
cores were collected. Eight Hexagenia nymphs (c.a, 40 mg/individual net 
weight) or 10 juvenile fathead minnows (c.a. 400 mg/individual net 
weight) were the test organisms. Mortality and biomass changes over 
three weeks were the endpoints examined. pH and dissolved oxygen were 
monitored in all chambers. 

In Site A, intact sediments resuUed in higher mortality and poorer 

growth than homogenized sediments for mayfly nymphs, but did not 
significantly influence mortality or growth in fathead minnows. Intact 
sediments from Site B resulted in better growth for mayfly nymphs than 
homogenized sediment. Mortality was <10% in both treatments. 
Homogeniration resulted in substantial mortality for fathead minnows 
(87X vs 20% in intact cores), in Site C (sandy sediment), 
homogenization resulted In higher mortality than in the intact cores for 
Hexagenia , This was most liKely caused by the elimination of the 
surface layer of fine-grained material (present in intact cores) and 
therefore, the elimination of suitable substrate for burial and feeding. 
Homogenization did not effect growth of fathead minnows, and may have 
ameliorated toxicity as measured by mortality. 


TABLE 3: Effect of Seitllng Time on Growth of Hexagenla 1 imbata 
Values in parentheses are standard deviations 





5h 24h 




Toronto STP 





Percent Biomass -3 
Change (Day 10) (0.9) 












Percent 4 
Mortality (Day 10) (5) 









Percent Biomass 4 
Change (Day 21) (9) 











Percent 7 

Mortal ity (Oay Zl) (9) 









As a result of these preliminary ocperiments, we recoTfimend further 
detailed examination of bioassay design and chronic endpoints, including 
bioaccumulation, in order to determine the significance of inplsce 
pol lutants. 


1. Lee. G.F., M.D. Piwoni, J.M. Lopez, G.M. Kariani. J.S. Richardson, 

O.H. Homer, and F. Saleh, 1975. Dredged Material Research 
Program, Contract Report 0-75-4, 

2. Ehuba, P.J., H.E, Tatem, and J.H. Carroll, 1978, Dredged Material 

Research Program Tech. Rep- O-78-50, 

3. U.S. Environmental Protection Agency/U.S. Army Corps of Engineers, 

1977. Waterways Experimental Station, Vicksburg, MS. 

4. Bahnick, O.A.. Swenson, W.A., Martiee. T.P., Call, D.J. Anderson, 

C.A,. and Morris, fi.T,, 1980. EPA Project No. R804918-C1. 

5. Swartz. R.C. D,W. Schults, E.R. Ditsworth and W,A. OeBen, 1984. 

Arch. Environ. Contara. Toxic. 13:207-216. 

6. Cairns, M.A., Nebeker, A.V., Gakstatter, J.H., and Griffis, W.L. 

(1984). Environ. Toxicol. Chem. 3:435-445, 




Sherrene D.Kevan and Dr. d.g. Dixon 

Department of Biology, University of Waterloo, Waterloo, Cnt. N2L 3Gl. 


Itiiocyanate (SO*-) is a pollutftnt of mining effluents when cyanidation is 
used to leach precious netal Erocn ores. Although SCN- effluents are released 
continuously, sfioct-term or pulse exposures of aquatic organisow occur because 
of accidencal spills, batch release of effluent, or organiama enter a mixing 

The iiT^MCt of SCN- pulse exposure on eggs of rainbcw; trout during their 
Aater hardening has not been investigated before. 

During water hardejiing, the diorion or fertilization mentorane of the egg 
becomes impervious to water penetrating into the egg. Before water hardening, 
for about one hour after the egg has been deposited, the egg rapidly absorbs 
v^acer from its surroundings. Thus, one would eiq^ect vnter soluble ard toxic 
ions, such as SCK- to ha'^e a awre pronounced effect in killing eggs before 
they water narden than afterwards. Tix aim of this study vbs to examine that 


Rainbow trout tsqgs weire obtained from Rainbow Springs Trout Farm 
(Thameaford, Ont.). 'IV» bioassays were conducted using 4 replicates, eac*i 
ODntaining lOQ to 150 eggs and 7 K5CN concentrations of (control), 90, 180, 
360, 720, 1440, 2880 rrg/1. The eggs were split into two groups. The first 
group «LS dry-f^rttlized and then exposed to KSCN [or 3 hours during water 


hardening. The aecorid group was fertilized and allowed to water harden in 
clean water for 3 hours before the 3 hour exposure to KSCN. Once the eggs 
were created, the/ were randomly assigned to canpartirents within B trays o£ a 
Plex-a-lite incubator where they were allowed Co develops up to hatch. 
During incubation, temperatures and flow rates of the water wera 
monitored. Fertility e.'Stimatea were made on subsamples of 50 egga preserved 
in StocAard'3 solution I4h postfertilization. Eggs in the incubator were 
checkod regularly and dead eggs reiDoved weekly. Egg rt»rtality and 
developmental abnormalities after hatch were recorded. Cni square analysis 
was carried out by ^tstat* program. The u:50 and 95% conEidence limits were 
determined by Trimmed Spearman-Karber analysis, 


The estimated LC50 for the treatment before hardening (iras 2018.749 (^S% 
confidence Uiuits = 1898.042 to 2H7.132) and for after hardening it was 
1530.268 1951 confidence limits » 1406.536 to 1662.520). Ihese are 
significantly different. 

Chi square analysis showed that there are no clearly significant 
differences in mortality. Before treatment (nortalities ranged from 16.8 to 
26. 0\ but after treatment mortalities ranged frcrn 19.3 to 23. 5S for 
concentrations cf 90 to 720 mg/l KSCN. At 1440 mg/1 there tes a highly 
significant differewe (p < 10''l. There «raa no significant difference in 
TOctalicy (78.8% before and 72.2% after) among treatments at the highest 
concentration 12880 mg/ll. Clearly, mortalities at 2880 mg/1 are significantly 
higher than at other ooocentrations tor both before and after hardening, Cwt 
at 1440 mg/1, only the eggs treated after hardening showed siOTiiflcanr 
difference from those treated with lower concentrations. 

Percent deformities were jreateat at 1440 and 2880 mg/1 in both 


treatments (ca. 2-6 X at lower cOTicentrationa, but between 7.1 * and 9.(, % 
at the higher concentrations). Fertilization rates v«re redifced in eggs 
treated both before and after hardening at 28B0 mg/l (by 5AS ar»d iOl 
respectively) versus at the Irat^r concentrations. 


Exposure to concentrations of KSCN had a more adverse effect on trout 
eggs after wator hardening for 3 hours than on eggs that had not vater- 
nardened. Thta result is contrary to the original hypothesis, but is 
consistent with the resales of other imrestigatocs . Perhaps the age and 
iietaboUc activity of eggs after hardening makes them more susceptible to 
poisoning. PertiUzation was affected at the highest concentrations in both 
treatments but was mare pronounced in the before hardening treatment. Eggs 
appear to be imre sensitve directly after fertilization. Deformities vrtiich 
were obser/ed in both treat-nentSy and at all concentratic»is, were not 
significantly different. The action of the .9CN- ion i5 affecting the 
nietabolic process of thf.s egg in before arA aft^c stages of hardening. Overall, 
figg^ of rainbow trout are most sensititi'/e to reductions in fertilizaticwi, 
deforroities in development, and mortality after water hardening has occurred 
than before. 





Oral Presentations 

A1 Science andPolicv; Photochemical Oxidants and Acid 
Bear ing Species K. L. De:Derjian. Atmospheric 
Science Research Center, State University of New 
York, Albany, New York. L'.S.A, 

A2 Relationship Between Forest Decline and Root Health 
inOntario Sugar Maple C. Adams. M. Egj'edandT. 
Hutch;nson'. Dept. of Botany, University of Toronto. 
Toronto. Ontario, 

A3 A Numerical Decline Index Rating System to Monitor 

Changes in Tree Condition of Hardwood Forest Species 

D. vcLaughlin-. w. Mcliveen. w. Gizyn. D. Corrigan, 
S- Pearson and R. Arnup, Air Resources Branch. 
Environment Ontario 

A4 Invest! gat ion of Short- term Mutagenicity and 

Chemical Composition of Organic Solvent Ex tractable 
Fraction of Coke OvenEmission A, J Horton*. N, 
Belson, K. Shaw and G. H. Thomas, Ontario Research 
Foundation, clarkson, Ontario 

AS Quantitative Measurements of the Genetic Effects of 
Inhaled Carcinogens in Pulmonary Fibroblasts are Not* 
passible J A. Heddle". A. BouchandJ.D. Gingerich. 
Dept. of Biology, York University. Downsview, 

AG Sensitivity of Asthma tic children to Air Pol lutioa: 

D Pengelly* and C Goldsmith, McMaster University. 
Hanilton. Ontario 

A7 Hazardous Contaminants inOntario: Environmental 
Fate and Human Exposure D Mackay*andS. Paterson, 
Institute? or Environniental Studies, University of 
Toronto, Toronto, Ontario 



Oral Presentations 

A8 Verification of the Cloud and Wet Deposition Fields 
of a MesoScale Model of Long-Range Transport of Air 
Pollutants H. R. Cho". S.T. SoongandJ.V, Irtbarne. 
Department of Physics, University of Toronto, 
Toronto, Ontario 

A9 Eulerian Model Evaluation M. Alvo. Department of 

Matheaatics, University of Ottawa. Ottawa, Ontario 

A10 Scale Model Studies and Development of Prediction 
Procedures for Heavy Gas Dispersion in Complex 
Terrain 1988 P. A- Irwin-, M-C. SturphyandK. C. 
Heidorn, Rowan Williams Davies and Irwin Inc. , 
Guelph. Ontario 

A1 1 An Investigation of Wind Genera ted Par tide 

Transport Rates within a Turbulent Boundary -Layer 

A. D. Ciccone", J.G, Kawall and J. F. Keffer, 
Department of Mechanical Engineering, University of 
Toronto, Toronto, Ontario 

A12 Incineration of Wastes K. Davies, Environmental 

Protection Off ice. City of Toronto, Toronto, Ontario 

A13 Detectabillty of step Trends in the Rate of 

Atmospheric Sulphate Deposition z.h. McBean*. M. G. 

KompterandG J. Farqiihar. Departaent of Civil 
Engineering. University of Waterloo. Waterloo. 

A14 Incinerator and Steel Plant Contributions to .Air 

Particulates as Determined by Size-Specific Receptor 
Modelling A.C. Chan", Z-J. ftangandR.E. Jervis. 
Dept. of Chemical Engineering, University of 
Toronto. Toronto. Ontario 




Oral Presentations 

A15 A study on the Sources of Ac id Precipitation in 

Ontario. Canada P.K. Hopke-andY. 2eng, Department 
of Civil Engineering, university of Illinois, 
U'rbana. Illinois, U. S. A. 

A16 Ad%'anced Techniques for Mobile Monitoring ofTrace 
Organics in Air G.B. DeBrou*. E, Singer, M, A. Sage, 
R. w. Bell. R.E. Chapman andD J. Ogner. Air Resources 
Branch, Environment Ontario 

A1 7 Atmospheric Trace Gas Measurements Using a Tunable 
Diode Laser Absorption SpectroDeter D.R. Hastie- 
andH. I. Schiff. Departaent of Chemistry, York 
University. Downsvie*, Ontario 

A18 Biomedical Waste Incineration Testing Program V. 

Ozvacic". G. Wong, C. .Warson. H. Clement. D. Rokosh, 
S. Suter, G. Horsneli, J.C. Hipfner, S. Burns andH. 
Corinthios. Environment Ontario 

A19 A study of High Temperature Photochemical Kinetics of 
Sulphur Dioxide andNitrogen Oxides For a Flue Gas 
Treatment Process J. Hunt", P. Fellin, K A Brice. D. 
Ernst, D. Glendenning and R. Caton, Concord 
Scientific. Toronto, andC. Fung and K. Smith. 
Environment Ontario 

A20 Modelling the Photochemical Decomposi tion of 

ChlorinatedPhenolsby Sunlight N.J. Bunce'andJ. S. 
Nakai. Dept. of Chemistry andBiochemistry. 
University of Guelph. Guelph. Ontario 




Poster Presentations 

AP1 stochastic Modelling of Dispersion from Single 

EievatedSources E, Robertson and P. J. Barry, Atomic 
Energy of Canada Limited, Chalk River Nuclear 
Laboratories, ChalkSiver, Ontario 

AP2 Feasibility study for Assessing and Modelling 

MlcroclimaCic Conditions on the Foothill Kame (Phase 
1) T.B. Shaw, Brock University, St. Catharines. 

APS Critical Evaluation of Atmospheric Pollutant 
Parameterizatlonfrom Satellite Imagery N.T. 
O'Neill, A. Royer and L. Hubert. Universitede 
Sherbrooke, Sherbrooke. Quebec. andJ. Miller andJ. 
Freemantle. CRESS. York University. 
Downsview . ■ .Ontario. andC. Austin and A. Davis. 

AP4 A 3-D Mesoscale Wind Field Model and its .Application 
for Emergency Planning at .Nuclear Power Plants in 
H. Sahota. P.K. Misra, R. BloxamandD. Ehee, Air 
Resources Branch, Environment Ontario 

APS The Results fromaMeso-scaleModel M. Niewiadomski, 

University of Toronto. Toronto. Ontario 

AP6 Dose Response for Selected Environmental Air 

Pollutants: Results from a Study onRunners R. B. 

L'rch. F. Silverman, P. Corey andR.J. Shephard. The 
Cage Research Institute, University of Toronto, 
Toronto. Ontario 



Poster Presentations 

APT HamiltonAlr: chemical ComposltionandGenotoxic 
Activity of Respirable Particulate and Organic 
Vapours D.W, Bryant, C. Kaiser-Farrell andD. R. 
McCalla, Department of Biochemistry, McMaster 
University. Hamilton. Ontario 

APS Hutageaiclty Studies andRlsk Estimation of Complex 
Mixtures of Organic AlrborneContaminants AS. Raj 

andD.M. Logan. Department of Biology. York 
University. Downsview, Ontario 

APS In-Situ Monitoring of the Environment for 

GenotoxicltyLevelsUsingRodents M. Petras. M. 
vrzoc. S. Meddins. K. HillandT- Sands. Department of 
Biological Sciences. University of Windsor, Windsor. 

API Method Development for the Moni toring and Analysis of 
Odorous Organics InAmblent Air C.C. Chan. L. Vainer 
andJ.w. Martin. MannTesting Laboratories Ltd. , 
Mississauga. Ontario, and A. Szakolcai andB. Foster. 
Environment Ontario 

AP11 Gas Phase .^alysls of Organic Compounds from 

Structural DomainModulationwlthinFlourescent 
LlpidMultllayers L-J Krull. R S Brown and K 
Stewart. Department of Chemistry, Erindale Cai:q>u3. 
University of Toronto, Mississauga. Ontario 

API 2 Atmospheric Measurements of Natural Hydrocarbons 

UsingGas Chromatography/Mass Spectrometry H. Niki 
anda.H. Khouw. Departaent of Chemistry and Centre 
for Atmospheric Chemistry. YorkUniversity. 
Downsview. Ontario 



Poster Presentations 

API 3 utilization of Established Air PollutionMoni torlng 
Networks in Ontario Following Nuclear Incidents 

J. A. SladeandG. Laszlo, Radiation and Industrial 
Safety Branch. Atomic Energj' of Canada Liaited. Chalk 
River. Ontario 

AP14ARe-Examlnation of Ontario's 24 Hour Ambient Air 

Quality Criterion for Hydrogen Fluoride R, D. Jones 
andD.S, Harper. ,\ir Resources Branch. Environjnent 

AP1 5 Product Ion of Ozone-insensitive White Bean Varieties 

T.E. Michaels, Department of Crop Science. 
University of Guelph. Guelph, Ontario 

API 6 Efficacy of Fi Im-formlng Chemicals for Protecting 

Roadside Trees AgainstSalt Spray C. Chong. Ministry 
of Agriculture andFood, Horticultural Research 
Institute of Ontario, Vineland Station, Ontario 

API 7 An Evaluation of the Problems of Particulate Emission 
from the WoodProducts Industry M. F. Lepage and A. E. 
Davies. RowanWilliamsDaviesi Irwin Inc. , Guelph. 

API 8 Relationship of Sugar Maple Decline and 

Corresponding Chemical Changes in Xylem Sap 
Carbohydrates, .^Ilcronutrients and Trace Element* 

S.N. Pathak, T. Hutchinson and D. N. Roy. Department 
of Forestry, University of Toronto. Toronto. Ontario 

AP19 Identlficatlonof Long Range Aerosol Sources at the 
DorsetEnvironment Station J. Drake, A. KabirandS, 
Vermette. Department of Geography, McMaster 
University. Hamilton, Ontario 




Oral Presentations 

CI An Overview of Hydrogeo logical Aspects of Waste 
Disposal: Research Results and Implications J. 

Cherry, Waterloo Centre for Groundwater Research, 
University of Waterloo, Waterloo, Ontario. 

C2 immiscibleHquids andVapours inSoil: Recent 

Experiments on Transport and Control G. Farquhar". 
R, Bensen, D. Graham, E- McBeanandB. Stickney, Dept- 
of Civil Sng. , University of Waterloo. Waterloo, 

C3 Effects of Increasing. Amounts of Non-polar Organic 
Liquids in Domestic Waste Leachate on the Hydraulic 
Conductivityof Clay Liners in Southern Ontario T. 

Fernandez- and a. M. Quigley. University of Western 
Ontario, London. Ontario. 

C4 Technology Review; Biological Treatment of Hazardous 
Landfill Leachates J. Fein'. andP- Yu, Diversified 
Research Laboratories Ltd- . Toronto, Ontario. 

C5 Phase Partitioning Kinetics at Industrial Waste Land 

Treatment Sites D. Hockley andW. J . Snodgrass-. Beak 
Consultants, Toronto. Ontario. 

C8 PreliminaryAssessment of aMicrofiltration/Reverse 
Osmosis Process for theTreatment of Landfill 
Leachate T.A. Krug"andS. McDougall. Zenon 
Environmental Inc. , Burlington. Ontario. 

C7 .inaerobicTreatment of Landf illLeachate G, P. 

vicevic-, B.J. Forrestal andA. Stevenson, Ontario 
ResearchFoundation. Clarkson. Ontario. 



Oral Presentations 

C8 The Origin and Distribution of Methane in the 

AllisconSandAquifer R. Aravena". J. Barker, M. 
Bliss andL. wassenaar. Department of EarthSciences, 
University of Waterloo. Waterloo. Ontario. 

C9 The carbon and sulfur cycle inShallowUnconf ined 
Aquifer Systems L. I. Wassenaar", B. Aravena. H, w, 
Gillbam. J. Barker andP. Fritz. Department of Earth 
Sciences- L'niversity of Waterloo. Waterloo, Ontario 

CIO Determination of Organic and Inorganic Contaminants 
in theWellandRiver ID, Brindle-, A.w, ChuandX-f 
Li, Chemistry Department, Brock University, St. 

Catharines, Ontario. 

C11 Research and Development of Permanent On-site 

Solutions for Contamination of Groundwater at Waste 
Disposal and Industrial Sites inCanada R J. Rush, 
CANVIROConsultants, Kitchener, Ontario. 

C12 The Roleof Groundwater inHunanSociety R.N. 

Farvolden. Waterloo Centre for Groundwater Research. 
University of Waterloo. Waterloo, Ontario. 

C13 Disperslonof theStouffvllleLandfillPlume I. 

Proulx* andfl. N". Farvolden, Waterloo Centre for 
GrouTidwater Hesearch, University of Waterloo. 

CI 4 comparison of an Experimental >runiclpal Refuse 

ColumnStudy withLandf ill FleldTestCella S. Pirani 
andD w Kirk-, Dept. oi' chenical Engineering, 
Univeraityof Toronto, Toronto. Ontario. 



Oral Presentations 

CI 5 An A Iter native to Incineration of Biamedical Vaste: 

HammeriLlll/Cheiaical Decontamination J. Manuel, 
Waste Management Branch, Environment Ontario. 

CI 6 Erosion of Landfill Covers J. Cuthill". Department 
of Land Resource Science. University of Guelph, 
Guelph, Ontario. andK. McKague, Ecoiogistics Ltd. , 
Waterloo, Ontario. 

CI 7 Development of Backfill and Construction Application 

Guidelines for Ontario M. Kelleher- andB, whiff in. 
C^-NViROConsuitants. Mississauga. Ontario. 

CI 8 Panal Discuasion: Stmming the Rising Tide 
of Hasta 

Moderator: D. Mackay, University of Toronto 




Poster Presentations 

CP1 Retractable Composite Absorbents for Envirotuoental 
Clean-up B. Gilliea. E. Stubley. 1, Treurnicht ar.d L 
Head, EcoPlastics Ltd, Willowdale. Ontario ando. 
Mereaz. Laboratory Services Branch. Environment 

CP2 Treatment and Disposal of Hauled Sewage Under ' , 'Part 
VII, Environmental ProtectionAct J.L, Smith, 
Oliver, Mangione, McCaliaiAssociates Limited, 
Nepean, Ontario 

CP3 Factors Affecting the Concentration of Metal Ions in 
Municipal Refuse Leachate C Kosta. S. PiraniandD 
Kirk. Department of Chemical Engineering and Applied 
Chemistry, University of Toronto, Toronto. Ontario. 

CP4 Slow Rate Infiltration Land Treatment and 

Recirculation of Landfill Leachate in Ontario R. A, 
.McBride, A.M. Gordon, P,H. Croenevelt, T.J. 
Gillespie andL. J. Evans, Departments of Land 
Resource Science and Environmental Biology, 
University of Guelph. Guelph, Ontario. 

CP5 Establishing Vegetation on Eros ion-prone Landfill 

Slopes inOntario. Year Two T.W. HilditchandC. P- 
Hughes, Gartner -Lee Ltd. , Marltham. Ontario, 

CP6 EvaluatlngGroundwater Velocity inaLow- 

PermeabllltyFractured Shale K.S. Novaitowski and 
J. A. Cherry. Centre for Groundwater Research. 
University of Waterloo, Waterloo, Ontario. 



Poster Presentations 

CP7 The Design and Evaluation of in-S ituBi ores torat ion 
Methods for the Treatment of Sludges and Soils at 
WasteDtsposal Sites K.L Berry-Spark and J. F. 
Parker. Centre for Groundwater Research. University 
of Waterloo. Waterloo, Ontario. 

CP8 Enhanced Biodegradat ion of .\romattc and Chlorinated 
Aliphatic Compounds In a Leachate- Impacted .iquifer 

■ D.W. Acton. M.Shaw, J.F, Barker, CI. .Mayf ield and 
J. A. Cherry, Univeristy of Waterloo, Waterloo, 

CP9 Waste Management Planning for Pharaaceutlcal 

Industry R. StarisandH. Makhija, Trent University. 
Peterborough, Ontario. 



Oral Presentations 

D1 Analytical chemistry inaRegulatoryEnvlronment R 

Kagel. Dow Chemicals, Midi ind. Michigan, U.S.A. 

D2 Adaptation of Water Preconcentr at ion Techniques 

Developed for PCDD.\na lysis to Other Target Organic 
Pollutants. E, Dowdall", B. B. Hollebone, L. 
Brownlee axidC. Shewchuk, Carleton University, 
Ottawa, Ontario, 

D3 The Purpose and Significance of Ultra trace .Analysis 
of Dibenzo-p-Dioxins: The Concept of Bisk L. 

Bro»-nlee" andB. R. Hollebone. Chemistry Department. 
Carleton University, Ottawa, Ontario. 

D4 Procedures for the.Uialysis of 2. 3. 7, 8 -Substituted 
PCDDiPCDFIsoners andOther Target Compounds In 
Environinental Samples F.W. Karasek*. T.S. Thompson 
andK.P. Naikwadi, University of Waterloo. Waterloo, 

D5 TheClosed-Loop StrippingTechnique, Applied to 

Potable Water to Solve Taste and Odour Problems J P. 
Palmentier*. D, Robinson and V. Taguchi, Laboratory 
Services Branch, Environment Ontajio. 

D6 Solid Supported Processes in Environmental Analysis 

J.M. Bosenfeld. Department of Pathoiogj-. McMaster 
University. Hamilton, Ontario. 

07 Synthesis and Use of Liquid Crystal! Ine Polys ilo»ane 
Substrate in Capillary Column GC -MS for Isomer 
Specific Separation of Toxic Isomers of PCDD and 
PCDF K.P Naikwadi" andF w. Karasek. Universityof 
Waterloo, Waterloo. Ontario. 



Oral Presentations 

D8 Development of Mobile Infrared Spectroscopy for On- 
site Spec iat ion of Organic Wastes P Yang' andj. 
Osborne, Laboratory Services 3ranch, Environment 

D9 Mobile Laboratory: On the Developmeot andlteal World 
Application Aspects D. Toner*. B. Dalton. D, Morse, 
K, Horn, P. YangandJ. Osborne, Laboratory Services 
Branch, E:nvironment Ontario. 

D10 Beglospec if ic Synthesis of All Isomeric 

NI trof luorenones and Ni trof luorenes by Trans i t ion 
Metal Catalyzed Cross Coup 1 ing Reactions V. 

Snieckus", T. lihama, J.-mFuandM. Bourguignon, 
University of Waterloo, Waterloo, Ontario. 

D1 1 Pr epar at ion of Heterocyclic Polynuclear Aromatic 
Compounds as .Analytical Standards E, Lee-Ruff', 
B. E. George. r.J. Ablenas and Y. S, Chung. Department 
of Chemistry, York University, Downsview, Ontario. 

D12 Application of ICP Spectrometry in Health and 
Environment: A Case Study of Soil Ingested by 
Children R. Barnes. University of Massachusetts, 

Amherst. Massachusetts, U.S.A. 

D13 Direct Sample Insertion Into an Inductively Coupled 
Plasma for Atomic Qniss ion and Mass Spectrometry L. 

Blain* andE.D. Salin, Department of Chemistry. 
McGill University, Montreal, Quebec. 



Oral Presentations 

D14 Analysis of Gersanium andTinby Hydride Gsneratlon 
D. C. Plasma Atomic Emission Spectrometry: 
Application toDetermioatioos of GertnaniumandTin in 
Air Filters I.D. Brindle", B. Buchanan and X-c. Le, 
Brock University. St. Catharines, Ontario. 

D15 Use of the Hot Slurry Technique for Solid Sample 

Introduction for ICP-AES L. Gervais* andE.D. Salin, 
Department of Chemistry. McCill University, 
Montreal. Quebec. 

D16 .advanced Technology for Destruction of Waterborne 
OrganicPollutanCs H, Al-Ekabi" andM Robertson. 
Nulite, A Division of Nut ech Energy System Inc. , 
London, Ontario. 

D1 7 Development of ACexperC 2: Implementation of an 

Expert System for Automated Metal Analysis by Atomic 
AbsorptionSpectroscopy M J. Stillman*. T.A. Cox 
andW.B. Browett, L'niversity of Western Ontario, 
London, Ontario. 

D18 Adap tat ion of Water Preconcentrat ion Techniques of 
Trace Metal Detection K, L Singfield*. B.B 
Hollebone, L.J. Brownlee. Dept. of 

Chemistry, Carleton University, Ottawa, Ontario, and 
P. Vijan, Environment Ontario. 

D13 Comparison of Various Leachate Extraction Procedures 
for the Characterization of Inorganics in Wastes 
J. R. Kramer", P. Brassard, J. deed and P,V, Collins, 
Department of Geology, McMaster University, 
Hamilton, Ontario. 



Oral Presentations 

D20 2, 4-DichIorophenoxylacetic Acid (2,4-D) 

Determination in Water . trine and Sol I Extracts by 
Enzyme Immunoassav (EIA) and Radioimmunoassay (BIA) 

J.C. Hall'andK. Krieg, Dept. of Environmental 
Biology, Universityof Guelph. Guelph, Ontai-io. 



Poster Presentations 

DPI Derlvatlzat ion of Acidic Organic Compounds Using 

Phase Transfer Catalysis V.Y Taguchi ando. w. Berg, 
Laboratory Services Branch, EnvironmentOntario, 

DP2 New Chemical Ionization Reagents Directed Toward 
Mass Spec trome trie. Analysis of Trace Organlcs T.B. 
McMahon, K. Froeseandc.E. Allison, Department of 
Chemistry and Guelph-waterloo Centre for Graduate 

Work inChemistry. L'niversity of ', 'Waterloo, 
Waterloo. Ontario. 

DP3 An InterruptedSegeoentedFlowStream 

Microwave' , -Solid Sample Decomposition for ICP-AES 

E.D. SalinAndB. Liu, Department of Chemistry, 
McGill L'niversity, Montreal, Quebec. 

DP4 Sol id Phase Ex tr action of P.\H's From Drinking Water 
and .Analysis of Chlorophenols andPhenoxy-acid 
Herbicides in water W.G. Craig and C. D. Hall. Paracel 
Laboratories Ltd. , Nepean, Ontario. 

DPS Automated Water Preconcentrat ion Sampler for Dioxin 
Detection at the Parts PerQuadrillionLevel C. 

Shewchuk. B. Kollebone, L. Brownlee andE. Dowdall. 
Carleton L'niversity. Ottawa. Ontario, andR. 
Hunsinger, M. Uza, H. TosineandS. Suter, 

DP6 .Automated KPIX Method for Low Level 

Polynuclear' . 'Aromatic Hydrocarbon (PAHl Analysis 
of Drinking Water P. w Crozier andC, D. Hall, 
Laboratory Services Branch. EnvironmentOntario. 



Poster Presentations 

DPT Supercritical Fluid Extraction of Trace Organics 
FromSolidMatricea P, KruusandR.C. Biirk. 
Departnent of Chemistry. Carle ton University. 
Ottawa. Ontario andC. Crawford, Laboratory Services 

Branch, Environment Ontario. 

DPS Automated Sample IntroductlonandFre- 

treatmenf , 'with Flow Injection ICP-ES J F- Hopper, 

F. MoandD.W. Boomer. Laboratory Services Branch. 
Environment Ontario, 

DPS Applications of Flow Injection Technologj" to ICP-MS 

M.J. Powell, J.F. Hopper andD. W. Boomer. Laboratory 
Services Branch, Eiivironment Ontario. 

DP10 Investigation of the In-Si tu .Ace ty la t ion Process and 
Its .Applicability to the Analysis of a Wide Range of 
Phenolic Compounds in Water R Lega, MereszandM. 
Savu. Laboratory Services Branch, Environment 

DP11 Robustness of the Student" sT- teat with Censored 
Environmental Quality Data E E. Creese, Creese 
Environmental Consulting, Waterloo. Ontario, 

DPI 2 Automation of Sol id SupportedReactlons by Robotics 
J-M. BosenfeldandE. Pevolinas, McMaster 
University, Hamilton. Ontario. 



Oral Presentations 

El Understanding Snvironmental-EconcDilc Integration 

P A. Victor, VHB Research Ltd. . Toronto, Ontario. 

E2 EconoDilc Valuation Disparities and Environmental 

Policies J.L. Knecsch. Economics Department. Simon 
FraserUniversity. Burnaby, British Columbia. 

E3 ThePhysco-social Impacts of Exposure to 

Environmental Contaminants in Ontario: A Feasibility 
Study S.M, Taylor*. J, Frank. M. Height. D. Strainer, 

S. Walter andN. White, McMaster University. 
Kanilton. Ontario, 

E4 Economic Assessments of MISA Regulations for Direct 
Industrial Dischargers in Ontario O.E. Salamon" and 
J, A. Donnan, Policy andPlaiuiing Branch, Environment 

E5 The Extra Strength Sewer Surcharge to Regulate 

Industrial Sanitary Waste Discharges M. Fortin", 
Ecologistics, Waterloo, Ontario, G. Zudovs. CANVIRO. 
andJ. Donnan andG, Zegarac, Environoent Ontario. 

EG A study of the Economic Factors Relating to the 
Implementation of Resource Recycl ing or Zero- 
Discharge Technologies for Heavy Metal Generating 
Industries In Canada B, Fleet*. J. Kassirer, T. 
Burreil, T. Sanger, C. SmallandB. Cardoza, 
University of Toronto, Toronto, Ontario, 

E7 Determinants of Participation in Sol id Waste Source- 
Separation Programs InHigh-Rise Apartment Buildings 

V.w. Maclaren. Department of Geography, University 
of Toronto, Toronto, Ontario. 



Poster Presentations 

EP1 The Sew Economics of Sustainable Development R, Z. 

Rivers, water Planning and Management Branch, Canada 
Centre For Inland waters, EnvironmentCanada, 
Burlington, Ontario. 

EP2 The Environmental Effects of Forestry Operations in 

Ontario: HowMuchDoWeKnow? J.A, Dunster, 
Federation of OntarioNaturalists. Toronto. Ontario. 


TD Environmental research 

172.5 technology transfer conference 

.057 ■'■^^^ proceedings / 

1988 20307 

vol. 2